Tomato variety NUN 00309 TOP

ABSTRACT

A new and distinct tomato variety NUN 00309 TOP is disclosed as well as seeds and plants and fruits thereof. NUN 00309 TOP is a processing tomato variety, comprising high resistance to Verticillium dahliae, Fusarium oxysporum f. sp. lycopersici Race 0, Race 1, and Race 2, and Tomato Spotted Wilt Virus, and intermediate resistance to Meloidogyne incognita.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority of U.S. Provisional Application No.62/966,733, filed on Jan. 28, 2020, which is hereby incorporated byreference in its entirety.

FIELD OF DISCLOSURE

The disclosure relates to the field of plant breeding and, morespecifically, to the tomato variety NUN 00309 TOP. The disclosurefurther relates to vegetative reproductions of tomato variety NUN 00309TOP, methods for tissue culture of tomato variety NUN 00309 TOP, methodsfor regenerating a plant from such a tissue culture, and to phenotypicvariants of tomato variety NUN 00309 TOP. The disclosure also relates toprogeny of tomato variety NUN 00309 TOP and the hybrid varietiesobtained by crossing tomato variety NUN 00309 TOP as a parent line withplants of other varieties or parent lines.

BACKGROUND

The goal of plant breeding is to combine various desirable traits in asingle variety. Such desirable traits may include greater yield,resistance to diseases, insects or other pests, tolerance to heat anddrought, better agronomic quality, higher nutritional value, enhancedgrowth rate and improved fruit properties.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: self-pollination andcross-pollination. A plant self-pollinates if pollen from one flower istransferred to the same or another flower of the same genotype. A plantcross-pollinates if pollen comes to it from a flower of a differentgenotype.

Plants that have been self-pollinated and selected for (uniform) typeover many generations become homozygous at almost all gene loci andproduce a uniform population of true breeding progeny of homozygousplants. A cross between two such homozygous plants of different linesproduces a uniform population of hybrid plants that are heterozygous formany gene loci. The extent of heterozygosity in the hybrid is a functionof the genetic distance between the parents. Conversely, a cross of twoplants each heterozygous at a number of loci produces a segregatingpopulation of hybrid plants that differ genetically and are not uniform.The resulting non-uniformity makes performance unpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants to makehybrids, and the evaluation of the hybrids resulting from the crosses.Pedigree breeding and recurrent selection are examples of breedingmethods that have been used to develop inbred plants from breedingpopulations. Those breeding methods combine the genetic backgrounds fromtwo or more plants or various other broad-based sources into breedingpools from which new lines are developed by selfing and selection ofdesired phenotypes. The new plants are evaluated to determine which havecommercial potential.

Tomato (Solanum lycopersicum and closely related species) is naturally adiploid and the basic chromosome number of the genus is x=12, most are2n=2x=24, including the cultivated ones. It originated in the New Worldand has since become a major food crop.

Tomato cultivars may be grouped by maturity, i.e., the time requiredfrom planting the seed to the stage where fruit harvest can occur.Standard maturity classifications include ‘early’, ‘midseason’ or‘late-maturing.’ Another classification for tomatoes is thedevelopmental timing of fruit set. ‘Determinate’ plants grow foliage,then transition into a reproductive phase of flower setting, pollinationand fruit development. Consequently, determinant cultivars have a largeproportion of the fruit ripen within a short time frame. Growers thatharvest only once in a season favor determinant type cultivars. Incontrast, ‘indeterminate’ types grow foliage, then enter a long phasewhere flower and fruit development proceed along with new foliar growth.Growers that harvest the same plants multiple times favor indeterminatetype cultivars.

Tomatoes can also be classified by their target markets: fresh marketand processing tomatoes. Fresh-market tomatoes are primarily used forsalads, salad bar and sandwiches, and require good storage properties.On the other hand, processing tomatoes generally requires red coloredand pink to red/crimson fruit flesh and higher percentage of solublesolids. Processing tomatoes can be canned whole, canned, diced orchopped, dried, roasted, pasted, pureed or concentrated, juiced, frozen,or put into ready-made dishes, for example, sauces, stews or soups.

In 2017, World Atlas (available at world-wide web word atlas underarticles/which-are-the-world-s-leading-tomato-producing-countries)reported that the worldwide production of tomatoes amounted to 170.8million tons. United States is ranked as the third largest producer oftomatoes in the world, next to China and India. Tomatoes are availablein the United States year-round, with California and Florida being themajor producers. Fresh-market tomatoes are available from May toDecember although supply peaks in July and in September through October.Processing tomatoes have the greatest supply from August to September.

In response to more recent consumer demands for dietary diversity,tomato breeders have developed a wider range of colors. In addition toexpanding the range of red colored fruits, there are cultivars thatproduce fruits that are creamy white, lime green, yellow, green, golden,orange and purple. Additionally, there are multi-colored varietiesexemplified by mainly red fruited varieties with green shoulders, andboth striped- and variegated-colored fruit.

SUMMARY OF VARIOUS ASPECTS OF THE DISCLOSURE

The disclosure provides for tomato variety NUN 00309 TOP, productsthereof, and methods of using the same. NUN 00309 TOP is a blocky, pearshaped, medium-sized processing tomato variety with thin viscosity, andextended field holding (EFH) capacity and is suitable for growing in theopen field.

In one aspect, the disclosure provides a seed of tomato variety NUN00309 TOP, wherein a representative sample of said seed has beendeposited under Accession Number NCIMB 44019. The disclosure alsoprovides for a plurality of seeds of tomato variety NUN 00309 TOP. Theseed of tomato variety NUN 00309 TOP may be provided as an essentiallyhomogeneous population of tomato seed. The population of seed of tomatovariety NUN 00309 TOP may be particularly defined as being essentiallyfree from other seed. The seed population may be grown into plants toprovide an essentially homogeneous population of tomato plants asdescribed herein.

In another aspect, the disclosure provides a plant or a progeny havingall the physiological and morphological characteristics of variety NUN00309 TOP when grown under the same environmental conditions. In anotheraspect, the plant or progeny has all or all but one, two, or three ofthe physiological and morphological characteristics of tomato varietyNUN 00309 TOP when measured under the same environmental conditions ande.g., evaluated at significance levels of 1%, 5% or 10% significance(which can also be expressed as a p-value) for quantitativecharacteristics and determined by type or degree for non-quantitativecharacteristics, wherein a representative sample of seed of variety NUN00309 TOP has been deposited under Accession Number NCIMB 44019. Inanother aspect, the plant or progeny has all or all but one, two, orthree of the physiological and morphological characteristics as listedin Tables 1-2 for variety NUN 00309 TOP when measured under the sameenvironmental conditions and e.g., evaluated at significance levels of1%, 5% or 10% significance (which can also be expressed as a p-value)for quantitative characteristics and determined by type or degree fornon-quantitative characteristics.

In another aspect, the plant of variety NUN 00309 TOP, or part thereof,or a progeny thereof has 14, 15, or more or all of the followingdistinguishing characteristics when compared to the Reference Variety asshown in Table 3: 1) nearly entire margin of leaflet; 2) no marginalrolling or wiltiness; 3) semi-erect leaf attitude in relation to mainstem; 4) larger size of leaflets; 5) semi-erect to horizontal attitudeof petiole of leaflet in relation to main axis; 6) longer mature leaf,7) wider mature leaf; 8) thicker petiole; 9) shorter peduncle; 10) pearmature fruit shape; 11) lighter intensity of green color excludingshoulder before maturity; 12) smaller peduncle scar; 13) larger core incross-section in relation to total diameter; 14) smaller stem scar; and15) resistant to Fusarium oxysporum f. sp. lycopersici Race 2, whendetermined at 5% significance level for numerical characteristics anddetermined by type or degree for non-numerical characteristics, whengrown under the same environmental conditions.

In another aspect, the plant of variety NUN 00309 TOP, or part thereof,or a progeny thereof comprises high resistance to Verticillium dahliae,Fusarium oxysporum f. sp. lycopersici Race 0, Race 1, and Race 2, andTomato Spotted Wilt Virus, and intermediate resistance to Meloidogyneincognita, measured according to UPOV standards described in TG/44/11.

The disclosure also provides a plant grown from a seed of tomato varietyNUN 00309 TOP and plant part thereof.

The disclosure furthermore provides a tomato fruit produced on a plantgrown from a seed of variety NUN 00309 TOP.

In another aspect, the disclosure provides a seed growing or grown on aplant of variety NUN 00309 TOP (i.e., produced after pollination of theflower of tomato variety NUN 00309 TOP).

In other aspects, the disclosure provides for a plant part obtained fromvariety NUN 00309 TOP, wherein said plant part is: a fruit, a harvestedfruit, a part of a fruit, a leaf, a part of a leaf, pollen, an ovule, acell, a petiole, a shoot or a part thereof, a stem or a part thereof, aroot or a part thereof, a root tip, a cutting, a seed, a part of a seed,seed coat or another maternal tissue which is part of a seed grown onsaid variety, a hypocotyl, a cotyledon, a scion, a stock, a rootstock, apistil, an anther, or a flower or a part thereof. Fruits areparticularly important plant parts. Such plant parts may be suitable forsexual reproduction, vegetative reproduction, or a tissue culture. Inanother aspect, the plant part obtained from variety NUN 00309 TOP is acell, optionally a cell in a cell or tissue culture. That cell may begrown into a plant of variety NUN 00309 TOP.

In another aspect, the disclosure provides for an inbred variety of NUN00309 TOP.

In another aspect, the disclosure provides for a hybrid tomato varietyNUN 00309 TOP. The disclosure also provides for a progeny of tomatovariety NUN 00309 TOP. In another aspect, the disclosure provides aplant or a progeny retaining all or all but one, two or three of the“distinguishing characteristics” or all or all but one, two, or three ofthe “morphological and physiological characteristics” of tomato varietyNUN 00309 TOP and methods for producing that plant or progeny.

The disclosure also provides a cell culture of tomato variety NUN 00309TOP and a plant regenerated from tomato variety NUN 00309 TOP, whereinthe plant has all the characteristics of variety NUN 00309 TOP, whengrown under the same environmental conditions, as well as methods forculturing and regenerating tomato variety NUN 00309 TOP. Alternatively,a regenerated plant may have one characteristic that is different fromtomato variety NUN 00309 TOP.

The disclosure further provides a vegetatively propagated plant ofvariety NUN 00309 TOP having all or all but one, two, or three of themorphological and physiological characteristics of tomato variety NUN00309 TOP, when grown under the same environmental conditions as well asmethods for vegetatively propagating tomato variety NUN 00309 TOP.

In another aspect, the disclosure provides a method of producing atomato plant comprising crossing a first parent tomato plant with asecond parent tomato plant and harvesting the resultant hybrid tomatoseed, wherein said first parent plant or second parent tomato plant istomato variety NUN 00309 TOP. Also provided is a hybrid tomato seedproduced from crossing a first parent tomato plant and second parenttomato plant and harvesting the resultant hybrid tomato seed, whereinsaid first parent plant or second parent tomato plant is tomato varietyNUN 00309 TOP. Moreover, the hybrid tomato plant grown from the hybridtomato seed is provided.

In another aspect, the disclosure provides a method of introducing asingle locus conversion into a plant of variety NUN 00309 TOP, wherein arepresentative sample of seed of tomato variety has been deposited underAccession Number NCIMB 44019, wherein the plant otherwise retains all ofthe physiological and morphological characteristics of tomato varietyNUN 00309 TOP and further comprises the single locus conversion.

In yet another aspect, the disclosure provides a method for introducinga desired trait into tomato variety NUN 00309 TOP, said method comprisestransforming the plant of variety NUN 00309 TOP with a transgene thatconfers the desired trait, wherein the transformed plant otherwise hasall of the physiological and morphological characteristics of tomatovariety NUN 00309 TOP and contains the desired trait.

The disclosure also provides a method of producing a modified tomatoplant with a desired trait, wherein the method comprises mutating theplant or plant part of tomato variety NUN 00309 TOP, wherein arepresentative sample of seed of said tomato variety has been depositedunder Accession Number NCIMB 44019, and wherein the mutated plantotherwise retains all of the physiological and morphologicalcharacteristics of tomato variety NUN 00309 TOP, and contains thedesired trait.

In one aspect, the single locus conversion or desired trait is yield,storage properties, color, flavor, size, firmness, fruit quality,enhanced nutritional quality, post-harvest quality, male sterility,herbicide tolerance, insect resistance, pest resistance, diseaseresistance, environmental stress tolerance, modified carbohydratemetabolism, modified protein metabolism, or ripening.

In another aspect, the disclosure provides a container comprising theplant, plant part, or seed of tomato variety NUN 00309 TOP.

Also provided is a food, a feed, or a processed product comprising aplant part of tomato variety NUN 00309 TOP, wherein the plant part is afruit or part thereof.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows the leaf of tomato variety NUN 00309 TOP.

FIG. 2 shows the leaf of the Reference Variety.

FIG. 3 shows the leaf comparison of tomato variety NUN 00309 TOP and theReference Variety.

FIG. 4 shows the green fruit cluster comparison of tomato variety NUN00309 TOP and the Reference Variety.

FIG. 5 shows the fruits of tomato variety NUN 00309 TOP.

FIG. 6 shows the fruits of the Reference Variety.

FIG. 7 shows the fruit comparison of tomato variety NUN 00309 TOP andthe Reference Variety.

FIG. 8 shows the fruit cross-section of tomato variety NUN 00309 TOP andthe Reference Variety.

FIG. 9 shows the stem scar diameter comparison of tomato variety NUN00309 TOP and the Reference Variety.

DEFINITIONS

“Tomato” refers herein to plants of the species Solanum lycopersicum, ora closely related species, and fruits thereof. Solanum lycopersicum isalso known as Lycopersicon lycopersicum (L.) H. Karst. or Lycopersiconesculentum Mill. The most commonly eaten part of a tomato is the fruitor berry.

“Cultivated tomato” refers to plants of Solanum lycopersicum, or aclosely related species (e.g., varieties, breeding lines or cultivars ofthe species S. lycopersicum as well as crossbreds thereof, or crossbredswith other Solanum species), cultivated by humans and having goodagronomic characteristics.

The terms “tomato plant designated NUN 00309 TOP,” “NUN 00309 TOP,” “NUN00309,” “NUN 00309 F1,” “00309 TOP,” “tomato 000309,” or “N 6473” areused interchangeably herein and refer to a tomato plant of variety NUN00309 TOP, representative sample of said seed is deposited underAccession Number NCIMB 44019.

A “seed of NUN 00309 TOP” refers to a tomato seed which can be growninto a plant of tomato variety NUN 00309 TOP, wherein a representativesample of viable seed is deposited under Accession Number NCIMB 44019. Aseed can be in any stage of maturity, for example, a mature, viableseed, or an immature, non-viable seed. A seed comprises an embryo andmaternal tissues.

An “embryo of NUN 00309 TOP” refers to an “F1 hybrid embryo” as presentin a seed of tomato variety NUN 00309 TOP, a representative sample ofsaid seed is deposited under Accession Number NCIMB 44019.

A “seed grown on NUN 00309 TOP” refers to a seed grown on a mature plantof variety NUN 00309 TOP or inside a fruit of tomato variety NUN 00309TOP. The “seed grown on NUN 00309 TOP” contains tissues and DNA of thematernal parent, tomato variety NUN 00309 TOP.

A “fruit of tomato variety NUN 00309 TOP” refers to a fruit containingmaternal tissues of tomato variety NUN 00309 TOP as deposited underAccession Number NCIMB 44019. The fruit comprises pericarp, septa,epidermis, columella, locular cavity, vascular bundles and optionallyseed. Pericarp, septa, epidermis, columella, locular cavity, vascularbundles, and seed coat of the seed are maternal tissues, e.g., they aregenetically identical to the plant on which they grow. In one aspect,the fruit contains seed grown on tomato variety NUN 00309 TOP. Inanother aspect, the fruit does not contain seed, i.e., the fruit isparthenocarpic. The skilled person is familiar with methods for inducingparthenocarpy. Those methods comprise chemically or genetically inducingparthenocarpy. Compounds suitable for chemically inducing parthenocarpycomprise auxins, gibberellins and cytokinins. Methods for geneticallyinducing parthenocarpy comprise the methods described in U.S. Pat. No.9,125,353, US 2002/0010953, U.S. Pat. No. 6,060,648, EP 1057401 and EP1428425, which are herein incorporated by reference in their entireties.

“Plant” includes the whole plant or any part or derivatives thereof,preferably having the same genetic makeup as the plant from which it isobtained.

“Plant part” includes any part of a plant, such as a plant organ (e.g.,harvested or non-harvested fruits), a plant cell, a plant protoplast, aplant cell tissue culture or a tissue culture from which a whole plantcan be regenerated, a plant cell that is intact in a plant, a clone, amicropropagation, plant callus, a plant cell clump, a plant transplant,a vegetative propagation, a seedling, a fruit, a harvested fruit, a partof a fruit, a leaf, a part of a leaf, pollen, an ovule, an embryo, apetiole, a shoot or a part thereof, a stem or a part thereof, a root ora part thereof, a root tip, a cutting, a seed, a part of a seed, ahypocotyl, a cotyledon, a scion, a graft, a stock, a rootstock, apistil, an anther, and a flower or parts of any of these and the like.Seed can be mature or immature. Pollen or ovules may be viable ornon-viable. Also, any developmental stage is included, such asseedlings, cuttings prior or after rooting, mature plants or leaves.Alternatively, a plant part may also include a plant seed whichcomprises one or two sets of chromosomes derived from the parent plant,e.g., from tomato variety NUN 00309 TOP. An F1 progeny produced fromself-pollination of tomato variety NUN 00309 TOP will thus comprise twosets of chromosomes derived from tomato variety NUN 00309 TOP, while anF1 progeny derived from cross-fertilization of tomato variety NUN 00309TOP will comprise only one set of chromosomes from tomato variety NUN00309 TOP and the other set of chromosomes from the other parent.

An “essentially homogeneous population of tomato seed” is a populationof seeds where at least 97%, 98%, 99% or more of the total population ofseed are seed of tomato variety NUN 00309 TOP.

An “essentially homogeneous population of tomato plants” is a populationof plants where at least 97%, 98%, 99% or more of the total populationof plants are plants of variety NUN 00309 TOP.

The phrase “essentially free from other seed” refers to a population ofseed where less than 3%, 2%, 1% or less of the total population of seedis seed that is not a tomato seed or, in another aspect, less than 3%,2%, 1% or less of the total population of seed is seed that is not seedof tomato variety NUN 00309 TOP.

“Harvest maturity” is referred to as the stage at which a tomato fruitis ripe or ready for harvest or the optimal time to harvest the fruitfor the market, for processing or for consumption. In one aspect,harvest maturity is the stage which allows proper completion of thenormal ripening.

“Harvested plant material” refers herein to plant parts (e.g., singlefruits or clusters of fruits detached from the whole plant), which havebeen collected for further storage and/or further use.

“Yield” means the total weight of all tomato fruits harvested perhectare of a particular line or variety. It is understood that “yield”expressed as weight of all tomato fruits harvested per hectare can beobtained by multiplying the number of plants per hectare times the“yield per plant”.

“Marketable yield” means the total weight of all marketable tomatofruits, especially fruit which is not cracked, damaged or diseased,harvested per hectare of a particular line or variety. A “marketablefruit” is a fruit that has commercial value.

“USDA descriptors” are the plant variety descriptors described fortomatoing the “Objective Description of Variety Tomato (Solanumlycopersicum or Lycopersicon esculentum Mill)”, as published by the USDepartment of Agriculture, Agricultural Marketing Service, Plant VarietyProtection Office, Beltsville, Md. 20705 and which can be downloadedfrom the world wide web at ams.usda.gov underservices/plant-variety-protection/pvpo-c-forms under tomato. “Non-USDAdescriptors” are other descriptors suitable for describing tomato.

“UPOV descriptors” are the plant variety descriptors described fortomato in the “Guidelines for the Conduct of Tests for Distinctness,Uniformity and Stability, TG/44/11 (Geneva 2011, revised 2018-10-30), aspublished by UPOV (International Union for the Protection of NewVarieties and Plants), and which can be downloaded from the world wideweb at upov.int/under_edocs/tgdocs/en/tg044.pdf and is hereinincorporated by reference in its entirety. Likewise, “UPOV methods” todetermine specific parameters for the characterization of tomato aredescribed at upov.int.

“Calibration book for Tomato (Lycopersicon esculentum Mill.)” refers tothe calibration book for tomato which provides guidance for describing atomato variety, as published by Naktuinbow (version 1, December 2010).The calibration book is based on the CPVO (Community Plant VarietyProtection Office) Protocol CPVO-TP/044/3 and UPOV TG/44/10.

“RHS” refers to the Royal Horticultural Society (RHS) which publishes anofficial botanical color chart quantitatively identifying colorsaccording to a defined numbering system. The chart may be purchased fromRoyal Horticulture Society Enterprise Ltd RHS Garden; Wisley, Woking;Surrey GU236QB, UK, e.g., the RHS color chart 2007.

“Reference Variety for NUN 00309 TOP” refers herein to variety AB 0311,a variety from Seminis, which has been planted in a trial together withNUN 00309 TOP. The characteristics of tomato variety NUN 00309 TOP arecompared with the characteristics of the Reference Variety as shown inTables 1-2. The distinguishing characteristics between tomato varietyNUN 00309 TOP and the Reference Variety are shown in Table 3.

A plant having “all the physiological and morphological characteristics”of a referred-to-plant means a plant showing the physiological andmorphological characteristics of the referred-to-plant when grown underthe same environmental conditions, preferably in the same experiment;the referred-to-plant can be a plant from which it was derived, e.g.,the progenitor plant, the parent, the recurrent parent, the plant usedfor tissue- or cell culture, etc. A physiological or morphologicalcharacteristic can be a numerical characteristic or a non-numericalcharacteristic. In one aspect, a plant has “all but one, two or three ofthe physiological and morphological characteristics” of areferred-to-plant, or “all the physiological and morphologicalcharacteristics” of Tables 1-3 or “all or all but one, two or three ofthe physiological and morphological characteristics” of Tables 1-3.

The physiological and/or morphological characteristics mentioned aboveare commonly evaluated at significance levels of 1%, 5% or 10% if theyare numerical (quantitative), or for having an identical degree (ortype) if not numerical (not quantitative), if measured under the sameenvironmental conditions. For example, a progeny plant or a Single LocusConverted plant or a mutated plant of variety NUN 00309 TOP, may haveone or more (or all) of the essential physiological and/or morphologicalcharacteristics of said variety listed in Tables 1-3, as determined atthe 5% significance level (i.e., p<0.05) for numerical characteristicsand determined by type or degree for non-numerical characteristics, whengrown under the same environmental conditions.

“Distinguishing characteristics” or “distinguishing morphological and/orphysiological characteristics” refers herein to the characteristicswhich distinguish (i.e., are different) between the new variety andother tomato varieties, such as the Reference Variety, when grown underthe same environmental conditions. The distinguishing characteristicsbetween tomato variety NUN 00309 TOP and Reference Variety are describedin Table 3. When comparing tomato variety NUN 00309 TOP with differentvarieties, the distinguishing characteristics may be different. In oneaspect, the distinguishing characteristics may therefore include atleast one, two, three or more (or all) of the characteristics listed inTables 1-2.

Tomato variety NUN 00309 TOP has the following distinguishingcharacteristics when compared to the Reference Variety as shown in Table3: 1) nearly entire margin of leaflet; 2) no marginal rolling orwiltiness; 3) semi-erect leaf attitude in relation to main stem; 4)larger size of leaflets; 5) semi-erect to horizontal attitude of petioleof leaflet in relation to main axis; 6) longer mature leaf; 7) widermature leaf, 8) thicker petiole; 9) shorter peduncle; 10) pear maturefruit shape; 11) lighter intensity of green color excluding shoulderbefore maturity; 12) smaller peduncle scar; 13) larger core incross-section in relation to total diameter; and 14) smaller stem scar;and 15) resistant to Fusarium oxysporum f. sp. lycopersici Race 2, whendetermined at 5% significance level for numerical characteristics anddetermined by type or degree for non-numerical characteristics whengrown under the same environmental conditions.

Thus, a tomato plant “comprising the distinguishing characteristics oftomato variety NUN 00309 TOP” (such as a progeny plant) refers herein toa plant which does not differ significantly from said variety in thedistinguishing characteristics above. Therefore, in one aspect, thedisclosure provides a plant that does not differ significantly fromtomato variety NUN 00309 TOP in the distinguishing characteristicsabove.

Similarity and differences between two different plant lines orvarieties can be determined by comparing the number of morphologicaland/or physiological characteristics (e.g., the characteristics aslisted in Tables 1-2) that are the same (i.e., statistically notsignificantly different) or that are different (i.e., statisticallysignificantly different) between the two plant lines or varieties whengrown under the same environmental conditions. A numericalcharacteristic is considered to be “the same” when the value for anumeric characteristic is not significantly different at the 1% (p<0.01)or 5% (p<0.05) significance level, using T-test, a standard method knownto the skilled person. A non-numerical or “degree” or “type”characteristic is considered “the same” when the values have the same“degree” or “type” when scored using USDA and/or UPOV descriptors, ifthe plants are grown under the same environmental conditions.

In one aspect, a statistical analysis of quantitative characteristicsshowing the degree of significance may be provided. Statisticalsignificance is the likelihood that a relationship between two or morevariables is caused by something other than chance, i.e., that thedifferences in the means for quantitative characteristics of tomatovariety NUN 00309 TOP and the Reference Variety are significant or dueto chance. For the purpose of proving differences or distinction betweentomato variety NUN 00309 TOP and the Reference Variety, a p-value of 5%(0.05) or lower is considered statistically significant. This means thatthere is only a 5% probability that the observed result could havehappened just by chance or random variation.

The statistical analysis is drawn from a small sample of at least 15plants or plant parts of tomato variety NUN 00309 TOP and the ReferenceVariety. Statistical points or parameters such as mean, minimum, median,maximum, and standard deviation are collected from the sample data toanalyze where the average is, how varied the data set is, and whetherthe data is skewed. For the purpose of determining whether the result ofa data set is statistically significant, a T-test Paired Sample Means isused, a statistical tool for proving significance of means of the twogroups (e.g., tomato variety NUN 00309 TOP and the Reference Variety) at5% significance level (p-value of 5% or 0.05).

“Variety” or “cultivar” means a plant grouping within a single botanicaltaxon of the lowest rank.

A “plant line” is, for example, a breeding line which can be used todevelop one or more varieties. A breeding line is typically highlyhomozygous.

“Harvested seeds” refer to seeds harvested from a line or variety, e.g.,produced after self-fertilization or cross-fertilization and collected.

“Hybrid variety” or “F1 hybrid” refers to the seeds harvested fromcrossing two inbred (nearly homozygous) parental lines. For example, thefemale parent is pollinated with pollen of the male parent to producehybrid (F1) seeds on the female parent.

“Locus” (plural loci) refers to the specific location, place or site ofa DNA sequence on a chromosome, where, for example, a gene or geneticmarker is found. A locus may confer a specific trait.

“Allele” refers to one or more alternative forms of a gene locus. All ofthese loci relate to one trait. Sometimes, different alleles can resultin different observable phenotypic traits, such as differentpigmentation. However, many variations at the genetic level result inlittle or no observable variation. If a multicellular organism has twosets of chromosomes, i.e., diploid, these chromosomes are referred to ashomologous chromosomes, i.e., diploid. Diploid organisms have one copyof each gene (and therefore one allele) on each chromosome. If bothalleles are the same, they are homozygotes. If the alleles aredifferent, they are heterozygotes.

“Genotype” refers to the genetic composition of a cell or organism.

“Phenotype” refers to the detectable characteristic of a plant, cell, ororganism, which characteristics are the manifestation of geneexpression.

“Haploid” refers to a cell or organism having one set of the two sets ofchromosomes in a diploid.

“Diploid” refers to a plant, a vegetative plant part(s), or seed which adiploid plant can be grown, having two sets of chromosomes, designatedherein as 2n.

“Triploid” refers to a plant, a vegetative plant part(s), or seed whicha diploid plant can be grown, having three sets of chromosomes,designated herein as 3n.

“Tetraploid” refers to a plant, a vegetative plant part(s), or seedwhich a diploid plant can be grown, having four sets of chromosomes,designated herein as 4n.

“Scion” refers to a part of the plant attached to the rootstock. Thisplant is selected for its stems, leaves, flowers, or fruits. The scioncontains the desired genes to be duplicated in future production by thestock/scion plant and may produce the desired tomato fruit.

“Rootstock” or “stock” refers to the plant selected for its roots, inparticular for the resistance of the roots to diseases or stress (e.g.,heat, cold, salinity etc.). Normally the quality of the fruit of theplant providing the rootstock is less important.

“Stock/scion” or grafted plant refers to a tomato plant comprising arootstock from one plant grafted to a scion from another plant.

“Grafting” refers to the method of joining of (genetically) differentplant parts, especially scions and rootstocks, together so that theygrow as a single plant. A grafted seedling or a grafted plant is aseedling or plant (produced by grafting) consisting of such differentplant parts and which grows as one plant.

The term “traditional breeding techniques” encompasses herein crossing,selfing, selection, doubled haploid production, embryo rescue,protoplast fusion, marker assisted selection, mutation breeding, etc. asknown to the breeder (e.g., methods other than geneticmodification/transformation/transgenic methods), by which, for example,a genetically heritable trait can be transferred from one tomato line orvariety to another.

“Tissue culture” or “cell culture” refers to a composition comprisingisolated cells of the same or a different type or a collection of suchcells organized into parts of a plant. Tissue culture of various tissuesof tomato and regeneration of plants therefrom is well known and widelypublished (see, e.g., Bhatia et al. (2004), Plant Cell, Tissue and OrganCulture 78: 1-21). Similarly, methods of preparing cell cultures areknown in the art.

“Regeneration” refers to the development of a plant from cell culture ortissue culture or vegetative propagation.

“Vegetative propagation,” “vegetative reproduction,” or “clonalpropagation” are used interchangeably herein and mean a method of takinga part of a plant and allowing that plant part to form at least roots,and also refer to the plant or plantlet obtained by that method.Optionally, the vegetative propagation is grown into a mature plant. Theskilled person is aware of what plant parts are suitable for use in themethod.

“Selfing” refers to self-pollination of a plant, i.e., the transfer ofpollen from the anther to the stigma of the same plant.

“Crossing” refers to the mating of two parent plants. The termencompasses “cross-pollination” and “selfing”.

“Cross-pollination” refers to the fertilization by the union of twogametes from different plants.

“Backcrossing” is a traditional breeding technique used to introduce atrait into a plant line or variety. The plant containing the trait iscalled the donor plant and the plant into which the trait is transferredis called the recurrent parent. An initial cross is made between thedonor parent and the recurrent parent to produce a progeny plant.Progeny plants which have the trait are then crossed to the recurrentparent. After several generations of backcrossing and/or selfing therecurrent parent comprises the trait of the donor. The plant generatedin this way may be referred to as a “single trait converted plant”. Thetechnique can also be used on a parental line of a hybrid.

“Progeny” as used herein refers to a plant obtained from a plantdesignated NUN 00309 TOP. A progeny may be obtained by regeneration ofcell culture or tissue culture or parts of a plant of said variety orselfing of a plant of said variety or by producing seeds of a plant ofsaid variety. In further aspects, progeny may also encompass plantsobtained from crossing of at least one plant of said variety withanother tomato plant of the same variety or another variety or(breeding) line, or with wild tomato plants. A progeny may comprise amutation or a transgene. A “first generation progeny” or is the progenydirectly derived from, obtained from, obtainable from or derivable fromthe parent plant by, e.g., traditional breeding methods (selfing and/orcross-pollinating) or regeneration (optimally combined withtransformation or mutation). Thus, a plant of variety NUN 00309 TOP isthe male parent, the female parent or both of a first generation progenyof that variety. Progeny may have all the physiological andmorphological characteristics of variety NUN 00309 TOP, when grown underthe same environmental conditions. Using common breeding methods such asbackcrossing or recurrent selection, mutation or transformation, one ormore specific characteristics may be introduced into said variety, toprovide a plant comprising all but 1, 2, or 3 or more of themorphological and physiological characteristics of variety NUN 00309 TOP(as listed in Tables 1-2).

The terms “gene converted” or “conversion plant” or “single locusconverted plant” in this context refer to tomato plants which aredeveloped by traditional breeding techniques e.g., backcrossing, or viagenetic engineering or through mutation breeding, wherein essentiallyall of the desired morphological and physiological characteristics ofthe parent variety or line are recovered, in addition to the one or moregenes transferred into the parent via e.g., the backcrossing technique(optionally including reverse breeding or reverse synthesis of breedinglines). It is understood that not only the addition of a furthercharacteristic (e.g., addition of gene conferring a furthercharacteristic, such as a disease resistance gene), but also thereplacement/modification of an existing characteristic by a differentcharacteristic is encompassed herein (e.g., mutant allele of a gene canmodify the phenotype of a characteristic).

Likewise, a “Single Locus Converted (Conversion) Plant” refers to plantsdeveloped by plant breeding techniques comprising or consisting ofmutation and/or by genetic transformation and/or by traditional breedingtechniques, such as backcrossing, wherein essentially all of the desiredmorphological and physiological characteristics of a tomato variety arerecovered in addition to the characteristics of the single locus havingbeen transferred into the variety via the above mentioned technique. Incase of a hybrid, the gene may be introduced in the male or femaleparental line.

“Transgene” or “chimeric gene” refers to a genetic locus comprising aDNA sequence which has been introduced into the genome of the plant bytransformation. A plant comprising a transgene stably integrated intoits genome is referred to as “transgenic plant.”

As used herein, the terms “resistance” and “tolerance” are usedinterchangeably to describe plants that show no symptoms orsignificantly reduced symptoms to a specified biotic pest, pathogen,abiotic influence or environmental condition compared to a susceptibleplant. These terms are optionally also used to describe plants showingsome symptoms but that are still able to produce marketable product withan acceptable yield.

“Average” refers herein to the arithmetic mean.

The term “mean” refers to the arithmetic mean of several measurements.The skilled person understands that the appearance of a plant depends tosome extent on the growing conditions of said plant. The mean, if notindicated otherwise within this application, refers to the arithmeticmean of measurements on at least 10 different, randomly selected plantsof a variety or line.

DETAILED DESCRIPTION OF VARIOUS ASPECTS OF THE DISCLOSURE

The disclosure also relates to the plant of variety NUN 00309 TOP,wherein a representative sample of seeds of said variety has beendeposited under the Budapest Treaty, with Accession number NCIMB 44019.NUN 00309 TOP is a blocky, pear shaped, medium-sized processing tomatovariety with thin viscosity, and extended field holding capacity (EFH)and is suitable for growing in the open field.

The disclosure further relates to tomato variety NUN 00309 TOP, whichwhen compared to the Reference Variety has the following distinguishingcharacteristics as shown in Table 3: 1) nearly entire margin of leaflet;2) no marginal rolling or wiltiness; 3) semi-erect leaf attitude inrelation to main stem; 4) larger size of leaflets; 5) semi-erect tohorizontal attitude of petiole of leaflet in relation to main axis; 6)longer mature leaf, 7) wider mature leaf, 8) thicker petiole; 9) shorterpeduncle; 10) pear mature fruit shape; 11) lighter intensity of greencolor excluding shoulder before maturity; 12) smaller peduncle scar; 13)larger core in cross-section in relation to total diameter; and 14)smaller stem scar; and 15) resistant to Fusarium oxysporum f. sp.lycopersici Race 2, when determined at 5% significance level fornumerical characteristics and determined by type or degree fornon-numerical characteristics, when grown under the same environmentalconditions. Also encompassed are parts of that plant.

In another aspect, the plant of variety NUN 00309 TOP, or part thereof,or a progeny plant thereof, comprises all of the following morphologicaland/or physiological characteristics (i.e., as indicated on the UPOVTest Guidelines for tomato) as shown in Tables 1-2, when grown under thesame environmental conditions. A part of this plant is also provided.

In another aspect, the plant of tomato variety NUN 00309 TOP, or partthereof, or a progeny thereof comprises high resistance to Verticilliumdahliae, Fusarium oxysporum f. sp. lycopersici Race 0, Race 1, and Race2, and Tomato Spotted Wilt Virus, and intermediate resistance toMeloidogyne incognita, measured according to UPOV standards described inTG/44/11.

The disclosure further provides a tomato plant which does not differfrom the physiological and morphological characteristics of the plant ofvariety NUN 00309 TOP as determined at the 1%, 2%, 3%, 4% or 5%significance level for numerical characteristics and determined by typeor degree for non-numerical characteristics, when grown under the sameenvironmental conditions. In a particular aspect, the plants aremeasured in the same trial (e.g., the trial is conducted as recommendedby USDA or UPOV). The disclosure also comprises a part of said plant,preferably a fruit or part thereof.

The morphological and/or physiological differences between two differentindividual plants described herein (e.g., between tomato variety NUN00309 TOP and a progeny of said variety) or between a plant of varietyNUN 00309 TOP, or progeny of said variety, or a plant having all, or allbut 1, 2, or 3 of the physiological and morphological characteristics oftomato variety NUN 00309 TOP (or all, or all but 1, 2, or 3 of thecharacteristics as listed in Tables 1-2) and another known variety caneasily be established by growing said variety next to each other or nextto the other variety (in the same field, under the same environmentalconditions), preferably in several locations which are suitable for saidtomato cultivation, and measuring morphological and/or physiologicalcharacteristics of a number of plants (e.g., to calculate an averagevalue and to determine the variation range/uniformity within thevariety). For example, trials can be carried out in Acampo Calif., USA(N 38 degrees 07′261″/W 121 degrees 18′ 807″, USA), whereby variouscharacteristics, for example, growth, form, plant habit, no. of nodesbetween inflorescence, onset of leaflet rolling, inflorescence, calyx,corolla color, fasciation, fruit shape, abscission layer, number oflocules, fruit surface, fruit color, flesh color, epidermis color,anthocyanin level, disease resistance, insect resistance, can bemeasured and directly compared for species of tomato.

Thus, the disclosure comprises tomato plant having one, two, or threephysiological and/or morphological characteristics which are differentfrom those of the plant of variety NUN 00309 TOP and which otherwise hasall the physiological and morphological characteristics of the plant ofvariety NUN 00309 TOP, e.g., when determined at the 5% significancelevel for numerical characteristics and determined by type or degree fornon-numerical characteristics when grown under the same environmentalconditions. In one aspect, the different characteristic(s) is/are resultof breeding with tomato variety NUN 00309 TOP and selection of a progenyplant comprising 1, 2, or 3 characteristics which are different than intomato variety NUN 00309 TOP. In another aspect, the differentcharacteristic is the result of a mutation (e.g., spontaneous mutationor a human induced mutation through e.g., targeted mutagenesis ortraditional mutagenesis such as chemically or radiation inducedmutagenesis), or it is the result of transformation.

The disclosure also relates to a seed of tomato variety NUN 00309 TOP,wherein a representative sample of said seed has been deposited underthe Budapest Treaty, with Accession number NCIMB 44019.

In another aspect, a seed of hybrid variety NUN 00309 TOP is obtainableby crossing the male parent of tomato variety NUN 00309 TOP, with thefemale parent of tomato variety NUN 00309 TOP, and harvesting the seedsproduced on the female parent. The resultant seeds of said variety canbe grown to produce plants of said variety.

In another aspect, a seed or a plurality of seeds of said variety arepackaged into a container of any size or type (e.g., bags, cartons,cans, etc.). The seed may be disinfected, primed and/or treated withvarious compounds, such as seed coatings or crop protection compounds.The seed produces the plant of variety NUN 00309 TOP.

In another aspect, the disclosure provides a plant grown from a seed oftomato variety NUN 00309 TOP and plant part thereof.

The disclosure also provides a tomato fruit produced on a plant grownfrom a seed of tomato variety NUN 00309 TOP.

In another aspect, the disclosure provides for a tomato plant part ofvariety NUN 00309 TOP, preferably a fruit or part thereof, arepresentative sample of seed from said variety is deposited under theBudapest Treaty, with Accession number NCIMB 44019.

Also provided is a plant of tomato variety NUN 00309 TOP, or a fruit orother plant part thereof, produced from a seed, wherein a representativesample of said seeds is deposited under the Budapest Treaty, withAccession Number NCIMB 44019.

Also provided is a plant part obtained from variety NUN 00309 TOP,wherein said plant part is a fruit, a harvested fruit, a part of afruit, a leaf, a part of a leaf, pollen, an ovule, a cell, a petiole, ashoot or a part thereof, a stem or a part thereof, a root or a partthereof, a root tip, a cutting, a seed, a part of a seed, seed coat oranother maternal tissue which is part of a seed grown on said variety, ahypocotyl, a cotyledon, a scion, a stock, a rootstock, a pistil, ananther, and a flower or a part thereof. Such plant parts may be suitablefor sexual reproduction (e.g., a pollen, a flower, an ovary, an ovule,an embryo, etc.), vegetative reproduction (e.g., a cutting, a root, astem, a cell, a protoplast, a leaf, a cotyledon, a meristem, etc.) ortissue culture (e.g., a leaf, a pollen, an embryo, a cotyledon, ahypocotyl, a cell, a root, a root tip, an anther, a flower, a seed, astem, etc.). Fruits are particularly important plant parts. Fruits maybe parthenocarpic, or seedless, or contain immature and/or nonviableseeds.

In a further aspect, the plant part obtained from variety NUN 00309 TOPis a cell, optionally a cell in a cell or tissue culture. That cell maybe grown into a plant of variety NUN 00309 TOP. A part of the plant ofvariety NUN 00309 TOP (or of progeny of that variety or of a planthaving all physiological and/or morphological characteristics but one,two or three which are different from those of tomato variety NUN 00309TOP) further encompasses any cells, tissues, or organs obtainable fromthe seedlings or plants in any stage of maturity.

The disclosure also provides a tissue or cell culture comprising cellsof tomato variety NUN 00309 TOP. Such a tissue culture can, for example,be grown on plates or in liquid culture, or be frozen for long termstorage. The cells of tomato variety NUN 00309 TOP used to start theculture can be selected from any plant part suitable for vegetativereproduction, or, in a particular aspect, can be cells of an embryo,meristem, a cotyledon, a hypocotyl, pollen, a leaf, an anther, a root, aroot tip, a pistil, a petiole, a flower, a fruit, a seed, or a stem. Inanother particular aspect, the tissue culture does not containsomaclonal variation or has reduced somaclonal variation. The skilledperson is familiar with methods to reduce or prevent somaclonalvariation, including regular reinitiation.

In one aspect, the disclosure provides a tomato plant regenerated fromthe tissue or cell culture of tomato variety NUN 00309 TOP, wherein theregenerated plant is not significantly different from tomato variety NUN00309 TOP, in all, or all but one, two, or three, of the physiologicaland morphological characteristics (e.g., determined at the 5%significance level when grown under the same environmental conditions).Optionally, the plant has one, two, or three the physiological andmorphological characteristics that is different from tomato variety NUN00309 TOP, wherein the difference or modification is effected bymutation or transformation with a transgene.

In another aspect, the disclosure provides a tomato plant regeneratedfrom the tissue or cell culture of tomato variety NUN 00309 TOP, whereinthe plant has all or all but one, two, or three of the physiological andmorphological characteristics of said variety determined (e.g., 5%significance level) when grown under the same environmental conditions.Similarity or difference of a characteristic is determined by measuringthe characteristics on a representative number of plants grown under thesame environmental conditions, determining whether type/degreecharacteristics are the same and determining whether numericalcharacteristics are different at the 5% significance level.

Tomato variety NUN 00309 TOP, or its progeny, or a plant having allphysiological and/or morphological characteristics but one, two, orthree which are different from those of tomato variety NUN 00309 TOP,can also be reproduced using vegetative reproduction methods. Therefore,the disclosure provides for a method of producing a plant or plant partof tomato variety NUN 00309 TOP, comprising vegetative propagation oftomato variety NUN 00309 TOP. Vegetative propagation comprisesregenerating a whole plant from a plant part of variety NUN 00309 TOP,from a progeny or from or a plant having all physiological and/ormorphological characteristics of said variety but one, two or threedifferent characteristics, such as a cutting, a cell culture, or atissue culture.

The disclosure also provides methods of vegetatively propagating apartof the plant of variety NUN 00309 TOP. In certain aspects, the methodcomprises: (a) collecting tissue or cells capable of being propagatedfrom tomato variety NUN 00309 TOP; (b) cultivating said tissue or cellsto obtain proliferated shoots; and (c) rooting said proliferated shoots,to obtain rooted plantlets. Steps (b) and (c) may also be reversed,i.e., first cultivating said tissue to obtain roots and then cultivatingthe tissue to obtain shoots, thereby obtaining rooted plantlets. Therooted plantlets may then be further grown, to obtain plants. In oneaspect, the method further comprises step (d) growing plants from saidrooted plantlets. Therefore, the method also comprises regenerating awhole plant from said part of the plant of variety NUN 00309 TOP. In aparticular aspect, the part of the plant to be propagated is a cutting,a cell culture, or a tissue culture.

The disclosure also provides for a vegetatively propagated plant ofvariety NUN 00309 TOP (or from progeny of tomato variety NUN 00309 TOPor from or a plant having all but one, two or three physiological and/ormorphological characteristics of that variety), wherein the plant hasall of the morphological and physiological characteristics of tomatovariety NUN 00309 TOP, when the characteristics are determined at the 5%significance level for plants grown under the same conditions. Inanother aspect, the propagated plant has all but one, two, or three ofthe morphological and physiological characteristics of tomato varietyNUN 00309 TOP, when the characteristics are determined at the 5%significance level for plants grown under the same conditions. A part ofsaid propagated plant or said propagated plant with one, two, or threedifferences is also provided. In another aspect, the propagated planthas all or all but 1, 2, or 3 of the physiological and morphologicalcharacteristics of tomato variety NUN 00309 TOP (e.g., as listed inTables 1-2).

In another aspect, the disclosure provides a method for producing atomato plant part, preferably a fruit, comprising: growing a plant ofvariety NUN 00309 TOP until it sets at least one fruit, and collectingthe fruit. Preferably, the fruit is collected at harvest maturity. Inanother aspect, the fruit is collected when the seed is ripe.

In another aspect, the plant of variety NUN 00309 TOP can be produced byseeding directly in the soil (e.g., field) or by germinating the seedsin controlled environment conditions (e.g., greenhouses, hydroponiccultures, etc.) and optionally then transplanting the seedlings into thefield. For example, the seed can be sown into prepared seed beds wherethey will remain for the entire production the crop (see, e.g., Hartz,et. al., University of California Division of Agriculture and NaturalResources, Publication 7228, 1-5). Tomatoes can be grown with a supportsystem such as poles (i.e., stakes) to keep the fruit from touching theground or as bushes without support. Alternatively, plastic row coverscan also be used to control the temperature. Mulches or plastic tunnelscan also be used to protect the plant from frost (see, e.g., Strange,et. al., 2000, University of California Division of Agriculture andNatural Resources, Publication 8017, 1-8). Tomato can also be grownentirely in greenhouses. Moreover, said variety can be grown inhydroponic cultures as described herein in, e.g., US 2008/0222949, whichis herein incorporated by reference in its entirety, and the skilledperson is familiar with various type of hydroponic cultures.

In another aspect, the plant and plant parts of tomato variety NUN 00309TOP and progeny of said variety are provided, e.g., grown from seeds,produced by sexual or vegetative reproduction, regenerated from theabove-described plant parts, or regenerated from cell or tissue cultureof the tomato variety NUN 00309 TOP, in which the reproduced (seedpropagated or vegetatively propagated) plant has all of thephysiological and morphological characteristics of tomato variety NUN00309 TOP, e.g., listed in Tables 1-2. In one aspect, said progeny oftomato variety NUN 00309 TOP can be modified in one, two, or threecharacteristics, in which the modification is a result of mutagenesis ortransformation with a transgene.

In other aspects, the disclosure provides a progeny plant of variety NUN00309 TOP, such as a progeny plant obtained by further breeding oftomato variety NUN 00309 TOP. Further breeding with tomato variety NUN00309 TOP includes selfing that variety one or more times and/orcross-pollinating tomato variety NUN 00309 TOP with another tomato plantor variety one or more times. In particular, the disclosure provides fora progeny plant that retains all the essential morphological andphysiological characteristics of tomato variety NUN 00309 TOP or, inanother aspect, a progeny plant that retains all, or all but one, two orthree, of the morphological and physiological characteristics of tomatovariety NUN 00309 TOP, optionally all or all but one, two, or three ofthe characteristics as listed in Tables 1-2, e.g., determined at the 5%significance level for numerical characteristics and determined by typeor degree for non-numerical characteristics when grown under the sameenvironmental conditions. In a particular aspect, the progeny is a firstgeneration progeny, i.e., the ovule or the pollen (or both) used in thecrossing is an ovule or pollen of variety NUN 00309 TOP, where thepollen comes from an anther and the ovule comes from an ovary of varietyNUN 00309 TOP. In another aspect, the disclosure provides for avegetative reproduction of the variety and a plant having all, or allbut 1, 2, or 3 of the physiological and morphological characteristics oftomato variety NUN 00309 TOP (e.g., as listed in Tables 1-2).

In still another aspect, the disclosure provides a method of producing atomato plant, comprising crossing a plant of tomato variety NUN 00309TOP with a second tomato plant at least once, allowing seed to developand optionally harvesting said progeny seed. The skilled person canselect progeny from said crossing. Optionally, the progeny (grown fromthe progeny seed) is crossed twice, thrice, or four, five, six, or seventimes, and allowed to set seed. In one aspect, the first “crossing”further comprises planting seeds of a first and a second parent tomatoplant, often in proximity so that pollination will occur; for example,mediated by insect vectors. Alternatively, pollen can be transferredmanually. Where the plant is self-pollinated, pollination may occurwithout the need for direct human intervention other than plantcultivation. After pollination the plant can produce seed.

The disclosure also provides a method for collecting pollen of tomatovariety NUN 00309 TOP, comprising collecting pollen from a plant ofvariety NUN 00309 TOP. Alternatively, the method comprises growing aplant of variety NUN 00309 TOP until at least one flower contains pollenand collecting the pollen. In a particular aspect, the pollen iscollected when it is mature or ripe. A suitable method for collectingpollen comprises collecting anthers or the part of the anther thatcontains pollen, for example, by cutting the anther or the part of theanther off. Pollen can be collected in a container. Optionally,collected pollen can be used to pollinate a tomato flower.

Instill another aspect, the disclosure provides a method of producing aplant, comprising selfing a plant of variety NUN 00309 TOP one or moretimes, and selecting a progeny tomato plant from said selfing. In oneaspect, the progeny plant retains all the distinguishing characteristicsof tomato variety NUN 00309 TOP, when grown under the same environmentalconditions. In a different aspect, the progeny plant comprises all ofthe physiological and morphological characteristic of tomato variety NUN00309 TOP of Tables 1-2.

The disclosure also provides a method for developing a tomato plant in atomato breeding program, using a tomato plant described herein, or itsparts as a source of plant breeding material. Suitable plant breedingtechniques are recurrent selection, backcrossing, pedigree breeding,mass selection, mutation breeding, and/or genetic marker enhancedselection. In one aspect, the method comprises crossing tomato varietyNUN 00309 TOP or progeny of said variety, or a plant comprising all but1, 2, or 3 or more of the morphological and physiologicalcharacteristics of tomato variety NUN 00309 TOP (e.g., as listed inTables 1-2), with a different tomato plant, and wherein one or moreoffspring of the crossing are subject to one or more plant breedingtechniques: recurrent selection, backcrossing, pedigree breeding, massselection, mutation breeding and genetic marker enhanced selection (see,e.g., Vidavsky and Czosnek, (1998) Phytopathology 88(9): 910-4). Forbreeding methods in general, see, e.g., Principles of Plant Genetics andBreeding, 2007, George Acquaah, Blackwell Publishing, ISBN-13:978-1-4051-3646-4.

In one aspect, pedigree selection is used as a breeding method fordeveloping a tomato variety. Pedigree selection is also known as the“Vilmorin System of Selection,” see, e.g., Allard, John Wiley & Sons,Inc., 1999, pp. 64-67. In general, selection is first practiced among F2plants. In the next season, the most desirable F3 lines are firstidentified, then desirable F3 plants within each line are selected. Thefollowing season and in all subsequent generations of inbreeding, themost desirable families are identified first, then desirable lineswithin the selected families are chosen. A family refers to lines thatwere derived from plants selected from the same progeny from thepreceding generation.

Thus, progeny in connection with pedigree selection are either thegeneration (seeds) produced from the first cross (F1) or selfing (S), orany further generation produced by crossing and/or selfing (F2, F3,etc.) and/or backcrossing (BC1, BC2, etc.) one or more selected plantsof the F1 and/or S1 and/or BC1 generation (or plants of any furthergeneration, e.g., F2) with another tomato plant (and/or with a wildrelative of tomato). Progeny may have all the physiological andmorphological characteristics of tomato variety NUN 00309 TOP, whengrown under the same environmental conditions and/or progeny may have(be selected for having) one or more of the distinguishingcharacteristics of tomato variety NUN 00309 TOP.

In yet a further aspect, the disclosure provides for a method ofproducing anew tomato plant. The method comprises crossing tomatovariety NUN 00309 TOP, or a plant comprising all but 1, 2, or 3 of themorphological and physiological characteristics of tomato variety NUN00309 TOP (as listed in Tables 1-2), or a progeny plant thereof, eitheras male or as female parent, with a second tomato plant (or a wildrelative of tomato) one or more times, and/or selfing a tomato plant ofvariety NUN 00309 TOP, or a progeny plant thereof, one or more times,and selecting progeny from said crossing and/or selfing. The secondtomato plant may, for example, be a line or variety of the speciesSolanum lycopersicon, S. chilense, S. habrochaites, S. pennelli, S.peruvianum, S. pimpinellifolium or other Solanum species.

In a further aspect, tomato variety NUN 00309 TOP is used in crosseswith other, different, tomato varieties to produce first generation (F1)tomato hybrid seeds and plants with superior characteristics. In aparticular aspect, the disclosure provides a tomato seed and a plantproduced by crossing a first parent tomato plant with a second parenttomato plant, wherein at least one of the first or second parent tomatoplant is tomato variety NUN 00309 TOP. In another aspect, the tomatoseed and plant produced are the first filial generation (F1) tomato seedand plants produced by crossing the plant of tomato variety NUN 00309TOP with another tomato plant.

The morphological and physiological characteristics (and thedistinguishing characteristics) of tomato variety NUN 00309 TOP areprovided in Tables 1-2, as collected in a trial according to USDA and/orUPOV standards. Encompassed herein is also a plant obtainable fromtomato variety NUN 00309 TOP e.g., by selfing and/or crossing and/orbackcrossing with said variety and/or progeny of said variety)comprising all or all but one, two or three of the physiological andmorphological characteristics of tomato variety NUN 00309 TOP listed inTables 1-2, e.g., as determined at the 5% significance level fornumerical characteristics and determined by type or degree fornon-numerical characteristics, when grown under the same environmentalconditions and/or comprising one or more (or all; or all except one, twoor three) characteristics when grown under the same environmentalconditions. The morphological and/or physiological characteristics mayvary somewhat with variation in the environment (such as temperature,light intensity, day length, humidity, soil, fertilizer use), which iswhy a comparison under the same environmental conditions is preferred.Colors can best be measured using the Royal Horticultural Society (RHS)Chart.

In another aspect, the disclosure provides a method of producing a plantderived from a tomato variety NUN 00309 TOP, comprising crossing a plantof variety NUN 00309 TOP either as a male or female parent with a secondplant or selfing tomato variety NUN 00309 TOP or vegetative reproductionof tomato variety NUN 00309 TOP and collecting seeds from said crossingor selfing or regenerating a whole plant from the vegetable cell-ortissue culture. Also provided are seeds and/or plants obtained by thismethod. All plants produced using tomato variety NUN 00309 TOP as aparent are within the scope of the disclosure including plant partsderived from tomato variety NUN 00309 TOP.

In further aspects, the method comprises growing a progeny plant of asubsequent generation and crossing the progeny plant of a subsequentgeneration with itself or a second plant and repeating the steps foradditional 3-10 generations to produce a plant derived from tomatovariety NUN 00309 TOP. The plant derived from tomato variety NUN 00309TOP may be an inbred line and the aforementioned repeating crossingsteps may be defined as comprising sufficient inbreeding to produce theinbred line. By selecting plants having one or more desirable traits ofthe line as well as potentially other selected traits.

The disclosure provides for methods of producing plants which retain allthe morphological and physiological characteristics of a plant describedherein. The disclosure also provides for methods of producing a plantcomprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of tomato variety NUN 00309 TOP (e.g., aslisted in Tables 1-2), but which are still genetically closely relatedto said variety. The relatedness can, for example, be determined byfingerprinting techniques (e.g., making use of isozyme markers and/ormolecular markers such as Single-nucleotide polymorphism (SNP) markers,amplified fragment length polymorphism (AFLP) markers, microsatellites,minisatellites, Random Amplified Polymorphic DNA (RAPD) markers,restriction fragment length polymorphism (RFLP) markers and others). Aplant is “closely related” to tomato variety NUN 00309 TOP if its DNAfingerprint is at least 80%, 90%, 95% or 98% identical to thefingerprint of that variety. In a particular aspect, AFLP markers areused for DNA fingerprinting (see, e.g., Vos et al. 1995, Nucleic AcidResearch 23: 4407-4414). A closely related plant may have a Jaccard'sSimilarity index of at least about 0.8, preferably at least about 0.9,0.95, 0.98 or more (see, e.g., Sharifova, S., et. al., (2013), Journalof Hort. Research, 21(1):83-89; Ince et al., (2010), Biochem. Genet.48:83-95; Parvathaneni et al., (2011), J. Crop Sci. Biotech, 14 (1):39-43; Pisanu, et. al., (2004), Acta Hort. 660, 83-89). The disclosurealso provides a plant and a variety obtained or selected by applyingthese methods on tomato variety NUN 00309 TOP. Such a plant may beproduced by crossing and/or selfing, or alternatively, a plant maysimply be identified and selected amongst plants of said variety, orprogeny of said variety, e.g. by identifying a variant within tomatovariety NUN 00309 TOP or within progeny of said variety (e.g., producedby selfing) which variant differs from the variety described herein inone, two or three of the morphological and/or physiologicalcharacteristics (e.g., in one, two or three distinguishingcharacteristics), e.g. those listed in Tables 1-2. In one aspect, thedisclosure provides a tomato plant having a Jaccard's Similarity indexwith tomato variety NUN 00309 TOP of at least 0.8, e.g. at least 0.85,0.9, 0.95, 0.98 or even at least 0.99.

In some aspects, the disclosure provides a tomato plant comprisinggenomic DNA having at least 95%, 96%, 97%, 98% or 99% sequence identitycompared to the genomic DNA sequence of a plant of variety NUN 00309 TOPas deposited under Accession Number NCIMB 44019. In some aspects, thetomato plant further comprises all or all but 1, 2, or 3 of themorphological and physiological characteristics of tomato variety NUN00309 TOP (e.g., as listed in Tables 1-2). In other aspects, the tomatoplant is a hybrid or other derived from a seed or plant of variety NUN00309 TOP. In other aspects, the tomato plant comprises thedistinguishing characteristics of tomato variety NUN 00309 TOP.

For the purpose of this disclosure, the “sequence identity” ofnucleotide sequences, expressed as a percentage, refers to the number ofpositions in the two optimally aligned sequences which have identicalresidues (×100) divided by the number of positions compared. A gap,i.e., a position in the pairwise alignment where a residue is present inone sequence but not in the other, is regarded as a position withnon-identical residues. A pairwise global sequence alignment of twonucleotide sequences is found by aligning the two sequences over theentire length according to the Needleman and Wunsch global alignmentalgorithm described in Needleman and Wunsch, 1970, J. Mol. Biol.48(3):443-53). A full implementation of the Needleman-Wunsch globalalignment algorithm is found in the needle program in The EuropeanMolecular Biology Open Software Suite (see, e.g., EMBOSS, Rice et al.,Trends in Genetics June 2000, vol. 16, No. 6. pp. 276-277).

In one aspect, a plant of variety NUN 00309 TOP may also be mutated (bye.g., irradiation, chemical mutagenesis, heat treatment, etc.) andmutated seeds or plants may be selected in order to change one or morecharacteristics of said variety. Methods such as TILLING (TargetingInduced Local Lesions in Genomes) may be applied to tomato populationsin order to identify mutants.

Similarly, tomato variety NUN 00309 TOP may be transformed andregenerated, whereby one or more chimeric genes are introduced into thevariety or into a plant comprising all but 1, 2, 3, or more of themorphological and physiological characteristics (e.g., as listed inTables 1-2). Many useful traits can be introduced into tomato varietyNUN 00309 TOP by e.g., crossing a tomato variety NUN 00309 TOP with atransgenic tomato plant comprising a desired transgene, as well as bydirectly introducing a transgene into tomato variety NUN 00309 TOP bygenetic transformation techniques.

Any pest or disease resistance genes may be introduced into a plant oftomato variety NUN 00309 TOP, progeny of said variety or into a plantcomprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of tomato variety NUN 00309 TOP (e.g., aslisted in Tables 1-2). Resistance to one or more of the followingdiseases or pests may be introduced into the plant described herein:Colorado potato beetle, Southern root knot nematode, Spider mites,Sugarfly beet army worm, Tobacco flea beetle, Tomato hornworm, Tomatofruitworm, Whitefly, Bacterial canker, Bacterial soft rot, Bacterialspeck, Bacterial wilt (Pseudomonas syringae pv. Tomato), Bacterial,Anthracnose (Gloeosporium piperatum), Brown rot or corky root(Pyrenochaeta lycopersici), Alternaria, Fusarium wilt (F. oxysporumraces), Gray leaf spot (Stemphylium spp.), Late blight (Phytophthorainfestans races), and Leaf mold (Cladosporium fulvum races), Nematode(Meloidogyne spp.), Verticillium Wilt (Verticillium dahliae), Ralstoniasolanacearum (Rs), Leveillula taurica (Lt), and/or Oidium neolycopersici(On). Other resistance genes, against pathogenic viruses (e.g., TomatoMosaic Virus (ToMV), Curly TOF Virus, Tomato Mottle Virus, Potato YVirus, Blotchey Ripening, Tobacco Etch Virus, the various Tobacco MosaicVirus races, Concentric cracking, Tomato Spotted Wilt Virus (TSWV),Tomato Yellow Leaf Curl Virus (TYLCV), Gold Fleck, Tomato Torrado Virus(ToTV)), fungi, bacteria, nematodes, insects or other pests may also beintroduced.

Genetic transformation may, therefore, be used to insert a selectedtransgene into the tomato plants of the disclosure described herein ormay, alternatively, be used for the preparation of transgenic tomatoplants which can be used as a source of the transgene(s), which can beintroduced into tomato variety NUN 00309 TOP by e.g., backcrossing. Agenetic trait which has been engineered into the genome of a particulartomato plant may then be moved into the genome of another tomato plant(e.g., another variety) using traditional breeding techniques which arewell-known in the art. For example, backcrossing is commonly used tomove a transgene from a transformed tomato variety into an alreadydeveloped tomato variety and the resulting backcross conversion plantwill then comprise the transgene(s).

Any DNA sequences, whether from a different species or from the samespecies, which are inserted into the genome using transformation, arereferred to herein collectively as “transgenes.” A “transgene” alsoencompasses antisense, or sense and antisense sequences capable of genesilencing. Thus, the disclosure also relates to transgenic plants oftomato variety NUN 00309 TOP. In some aspects, a transgenic plant oftomato variety NUN 00309 TOP may contain at least one transgene butcould also contain at least 1, 2, 3, 4, or more transgenes.

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to aregulatory element active in plant cells (e.g., promoter). Theexpression vector may contain one or more such operably linkedgene/regulatory element combinations. The vector may be in the form of aplasmid and can be used alone or in combination with other plasmids toprovide transformed tomato plants using transformation methods toincorporate transgenes into the genetic material of the tomato plant(s).Transformation can be carried out using standard methods, such asAgrobacterium tumefaciens mediated transformation, electroporation,biolistics particle delivery system, or microprojectile bombardment,followed by selection of the transformed cells and regeneration intoplants.

Plants can also be genetically engineered, modified, or manipulated toexpress various phenotypes of horticultural interest. Through thetransformation of tomato, the expression of genes can be altered toenhance disease resistance, insect resistance, herbicide resistance,stress tolerance, horticultural quality, and other traits.Transformation can also be used to insert DNA sequences which control orhelp control male sterility or fertility restoration. DNA sequencesnative to tomato as well as non-native DNA sequences can be transformedinto tomato and used to alter levels of native or non-native proteins.Various promoters, targeting sequences, enhancing sequences, and otherDNA sequences can be inserted into the genome for the purpose ofaltering the expression of proteins. Reduction of the specific activityof specific genes (also known as gene silencing or gene suppression) isdesirable for several aspects of genetic engineering in plants.

Genome editing is another method recently developed to geneticallyengineer plants. Specific modification of chromosomal loci or targetedmutation can be done through sequence-specific nucleases (SSNs) byintroducing a targeted DNA double strand break in the locus to bealtered. Examples of SSNs that have been applied to plants are: fingernucleases (ZFNs), transcription activator-like effector nucleases(TALENs), engineered homing endonucleases or meganucleases, andclustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 (Cas9), see, e.g., Songstad, et.al., Critical Reviews in Plant Sciences, 2017, 36:1, 1-23.

Thus, the disclosure also provides a method of producing a tomato planthaving a desired trait comprising mutating the plant or plant part ofvariety NUN 00309 TOP and selecting a plant comprising the desiredtrait, wherein the mutated plant retains all or all but one of thephysiological and morphological characteristics of variety NUN 00309TOP, optionally as described in Tables 1-2, and contains the desiredtrait and wherein a representative sample of seed of variety NUN 00309TOP is deposited under Accession Number NCIMB 44019. In a furtheraspect, the desired trait is yield, storage properties, color, flavor,size, firmness, fruit quality, enhanced nutritional quality,post-harvest quality, male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, environmental stresstolerance, modified carbohydrate metabolism, modified protein metabolismor ripening, or the mutation occurs in any of the following genes acs2,acs4, rin, pp2c1, arf9, intense, myb12.

The disclosure also provides a method for inducing a mutation in tomatovariety NUN 00309 TOP comprising:

-   -   a) exposing the seed, plant, plant part, or cell of tomato        variety NUN 00309 TOP to a mutagenic compound or to radiation,        wherein a representative sample of seed of said tomato variety        is deposited under Accession Number NCIMB 44019;    -   b) selecting the seed, plant, plant part, or cell of tomato        variety NUN 00309 TOP having a mutation; and    -   c) optionally growing and/or multiplying the seed, plant, plant        part, or cell of tomato variety NUN 00309 TOP having the        mutation.

The disclosure also provides a method of producing a tomato plant havinga desired trait, wherein the method comprises transforming the tomatoplant with a transgene that confers the desired trait, wherein thetransformed plant otherwise retains all of the physiological andmorphological characteristics of the plant of variety NUN 00309 TOP Fand contains the desired trait. Thus, a transgenic tomato plant isprovided which is produced by the method described above, wherein theplant otherwise has all of the physiological and morphologicalcharacteristics of tomato variety NUN 00309 TOP and the desired trait.

In another aspect, the disclosure provides a method of producing aprogeny of plant of variety NUN 00309 TOP, further comprising a desiredtrait, said method comprising transforming the plant of tomato varietyNUN 00309 TOP, with at least one transgene that confers the desiredtrait and/or crossing the plant of tomato variety NUN 00309 TOP with atransgenic tomato plant comprising a desired transgene so that thegenetic material of the progeny that resulted from the cross containsthe desired transgene(s). Also encompassed is the progeny produced bythis method.

A desired trait (e.g., gene(s) conferring pest or disease resistance, ortolerance for protection, etc.) can be introduced into tomato varietyNUN 00309 TOP, or progeny of said variety, by transforming said varietyor progeny of said variety with a transgene that confers the desiredtrait, wherein the transformed plant retains all or all but one, two orthree of the morphological and/or physiological characteristics ofvariety NUN 00309 TOP and contains the desired trait. In another aspect,the transformation or mutation confers a trait wherein the trait isyield, storage properties, color, flavor, size, firmness, fruit quality,enhanced nutritional quality, post-harvest quality, male sterility,herbicide tolerance, insect resistance, pest resistance, diseaseresistance, environmental stress tolerance, modified carbohydratemetabolism, modified protein metabolism or occurs in the intense gene.In a particular aspect, the specific transgene may be any known in theart or listed herein, including, a polynucleotide sequence conferringresistance to imidazolinone, sulfonylurea, glyphosate, glufosinate,triazine, benzonitrile, cyclohexanedione, phenoxy proprionic acid andL-phosphinothricin or a polynucleotide conferring resistance to Coloradopotato beetle, Southern root knot nematode, Spider mites, Sugarfly beetarmy worm, Tobacco flea beetle, Tomato hornworm, Tomato fruitworm,Whitefly, Bacterial canker, Bacterial soft rot, Bacterial speck,Bacterial wilt (Pseudomonas syringae pv. Tomato), Bacterial, Anthracnose(Gloeosporium piperatum), Brown rot or corky root (Pyrenochaetalycopersici), Alternaria, Fusarium wilt (F. oxysporum races), Gray leafspot (Stemphylium spp.), Late blight (Phytophthora infestans races), andLeaf mold (Cladosporium fulvum races), Nematode (Meloidogyne spp.),Verticillium Wilt (Verticillium dahliae), Ralstonia solanacearum (Rs),Leveillula taurica (Lt), and/or Oidium neolycopersici (On). Otherresistance genes, against pathogenic viruses (e.g., Tomato Mosaic Virus(ToMV), Curly TOF Virus, Tomato Mottle Virus, Potato Y Virus, BlotcheyRipening, Tobacco Etch Virus, the various Tobacco Mosaic Virus races,Concentric cracking, Tomato Spotted Wilt Virus (TSWV), Tomato YellowLeaf Curl Virus (TYLCV), Gold Fleck, Tomato Torrado Virus (ToTV)),fungi, bacteria, nematodes, insects or other pests may also beintroduced.

By crossing and/or selfing, (one or more) single traits may beintroduced into tomato variety NUN 00309 TOP (e.g., using backcrossingbreeding schemes), while retaining the remaining morphological andphysiological characteristics of said variety and/or while retaining oneor more or all distinguishing characteristics. A single trait convertedplant may thereby be produced. For example, disease resistance genes maybe introduced, genes responsible for one or more quality traits, yield,etc. Both single genes (e.g., dominant or recessive) and one or moreQTLs (quantitative trait loci) may be transferred into tomato varietyNUN 00309 TOP by breeding with said variety.

Alternatively, a single trait converted plant or single locus convertedplant of variety NUN 00309 TOP may be produced by (i) geneticallytransforming or mutating cells of tomato variety NUN 00309 TOP; (ii)growing the cells into a plant; and (iii) optionally selecting a plantthat contains the desired single locus conversion. The skilled person isfamiliar with various techniques for genetically transforming a singlelocus in a plant cell, or mutating said cells.

In another aspect, the disclosure provides a method of introducing asingle locus conversion, single trait conversion, or a desired traitinto tomato variety NUN 00309 TOP, comprising introducing a single locusconversion, single trait conversion, or a desired trait in at least oneof the parents of tomato variety NUN 00309 TOP; and crossing theconverted parent with the other parent of tomato variety NUN 00309 TOP,to obtain seed of said variety.

In another aspect, the step of introducing a single locus conversion,single trait conversion, or a desired trait in at least one of theparents comprises:

-   -   a) crossing the parental line of tomato variety NUN 00309 TOP        with a second tomato plant comprising the single locus        conversion, the single trait conversion, or the desired trait;    -   b) selecting F1 progeny plants that contain the single locus        conversion, the single trait conversion, or the desired trait;    -   c) crossing said selected progeny plants of step b) with the        parental line of step a) to produce a backcross progeny plant;    -   d) selecting backcross progeny plants comprising the single        locus conversion, the single trait conversion or the desired        trait and otherwise all or all but one, two or three of the        morphological and physiological characteristics the parental        line of step a) to produce selected backcross progeny plants;        and    -   e) optionally repeating steps c) and d) one or more times in        succession to produce selected second, third or fourth or higher        backcross progeny plants comprising the single locus conversion,        the single trait conversion or the desired trait and otherwise        all or all but one, two or three of the morphological and        physiological characteristics the parental line of step a) to        produce selected backcross progeny plants, when grown in the        same environmental conditions.        The disclosure further relates to plants obtained by this        method.

In another aspect, the step of introducing a single locus conversion,single trait conversion, or a desired trait in at least one of theparents comprises:

-   -   a) obtaining a cell or tissue culture of cells of the parental        line of tomato variety NUN 00309 TOP;    -   b) genetically transforming or mutating said cells;    -   c) growing the cells into a plant; and    -   d) optionally selecting plants that contain the single locus        conversion, the single trait conversion, or the desired trait.

In another aspect, the disclosure provides a method of introducing asingle locus conversion, single trait conversion, or a desired traitinto tomato variety NUN 00309 TOP comprising:

-   -   a) obtaining a combination of a parental lines of tomato variety        NUN 00309 TOP, optionally through reverse synthesis of breeding        lines,    -   b) introducing a single locus conversion, single trait        conversion, or a desired trait in at least one of the parents of        step a); and    -   c) crossing the converted parent with the other parent of        step a) to obtain seed of tomato variety NUN 00309 TOP.

In another method, the step of introducing a single locus conversion,single trait conversion, or a desired trait in at least one of theparents comprises genetically transforming or mutating cells theparental line of tomato variety NUN 00309 TOP; growing the cells into aplant; and optionally selecting plants that contain the single locusconversion, the single trait conversion, or the desired trait.

In any of the above methods, where the single locus conversion, singletrait conversion, or a desired trait concerns a trait, the trait may beyield or pest resistance or disease resistance. In one aspect, the traitis disease resistance and the resistance are conferred to Coloradopotato beetle, Southern root knot nematode, Spider mites, Sugarfly beetarmy worm, Tobacco flea beetle, Tomato hornworm, Tomato fruitworm,Whitefly, Bacterial canker, Bacterial soft rot, Bacterial speck,Bacterial wilt (Pseudomonas syringae pv. Tomato), Bacterial, Anthracnose(Gloeosporium piperatum), Brown rot or corky root (Pyrenochaetalycopersici), Alternaria, Fusarium wilt (F. oxysporum races), Gray leafspot (Stemphylium spp.), Late blight (Phytophthora infestans races), andLeaf mold (Cladosporium fulvum races), Nematode (Meloidogyne spp.),Verticillium Wilt (Verticillium dahliae), Ralstonia solanacearum (Rs),Leveillula taurica (Lt), and/or Oidium neolycopersici (On). Otherresistance genes, against pathogenic viruses (e.g., Tomato Mosaic Virus(ToMV), Curly TOF Virus, Tomato Mottle Virus, Potato Y Virus, BlotcheyRipening, Tobacco Etch Virus, the various Tobacco Mosaic Virus races,Concentric cracking, Tomato Spotted Wilt Virus (TSWV), Tomato YellowLeaf Curl Virus (TYLCV), Gold Fleck, Tomato Torrado Virus (ToTV)),fungi, bacteria, nematodes, insects or other pests may also beintroduced.

The disclosure also provides a plant having one, two or threephysiological and/or morphological characteristics which are differentfrom those of tomato variety NUN 00309 TOP, and which otherwise has allthe physiological and morphological characteristics of said variety,wherein a representative sample of seed of tomato variety NUN 00309 TOPis deposited under Accession Number NCIMB 44019. In particular, variantswhich differ from tomato variety NUN 00309 TOP, in none, one, two orthree of the characteristics mentioned in Tables 1-2 are encompassed.

The disclosure also provides a tomato plant comprising at least a firstset of the chromosomes of tomato variety NUN 00309 TOP, a sample of seedof said variety is deposited under Accession Number NCIMB 44019;optionally further comprising a single locus conversion or a mutation,wherein said plant has essentially all of the morphological andphysiological characteristics of the plant comprising at least a firstset of the chromosomes of said variety. In another aspect, this singlelocus conversion confers a trait, wherein the trait is yield, storageproperties, color, flavor, male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, environmental stresstolerance, modified carbohydrate metabolism, modified protein metabolismor ripening, or the mutation occurs in any of the following genes acs2,acs4, rin, pp2c1, arf9, intense, myb12.

In one aspect, the disclosure provides for a haploid plant and/or adoubled haploid plant of tomato variety NUN 00309 TOP or of a planthaving all but one, two or three physiological and/or morphologicalcharacteristics of tomato variety NUN 00309 TOP, or progeny of any ofthese, is encompassed herein. Haploid and doubled haploid (DH) plantscan, for example, be produced by cell or tissue culture and chromosomedoubling agents and regeneration into a whole plant. DH productionchromosome doubling may be induced using known methods, such ascolchicine treatment or the like. In one aspect, the method comprisesinducing a cell or tissue culture with a chromosome doubling agent andregenerating the cells or tissues into a whole plant.

In another aspect, the disclosure comprises a method for making doubledhaploid cells of tomato variety NUN 00309 TOP, comprising making doubledhaploid cells from haploid cells from the plant or plant part of tomatovariety NUN 00309 TOP with a chromosome doubling agent, such ascolchicine treatment (see, e.g., Nikolova V, Niemirowicz-Szczytt K(1996) Acta Soc Bot Pol 65:311-317).

In another aspect, the disclosure provides for haploid plants and/ordoubled haploid plants derived from tomato variety NUN 00309 TOP that,when combined, make a set of parents of tomato variety NUN 00309 TOP.The haploid plant and/or the doubled haploid plant of variety NUN 00309TOP can be used in a method for generating parental lines of tomatovariety NUN 00309 TOP.

The disclosure also provides methods for determining the identity ofparental lines of plants described herein, in particular the identity ofthe female line. US 2015/0126380, which is hereby incorporated byreference, relates to a non-destructive method for analyzing maternalDNA of a seed. In this method, the DNA is dislodged from the seed coatsurface and can be used to collect information on the genome of thematernal parent of the seed. This method for analyzing maternal DNA of aseed comprises contacting a seed with a fluid to dislodge DNA from theseed coat surface, and analyzing the DNA thus dislodged from the seedcoat surface using methods known in the art. The skilled person is thusable to determine whether a seed has grown on a plant of variety NUN00309 TOP or is a progeny of said variety, because the seed coat of theseed is a maternal tissue genetically identical to tomato variety NUN00309 TOP. In one aspect, the present disclosure relates to a seed coatcomprising maternal tissue of tomato variety NUN 00309 TOP. In anotheraspect, the disclosure relates to a tomato seed comprising a maternaltissue of tomato variety NUN 00309 TOP. In another particular aspect,the disclosure provides for a method of identifying the female parentalline of tomato variety NUN 00309 TOP by analyzing the seed coat of aseed of that variety. In another aspect, the disclosure provides for amethod of determining whether a seed is grown on tomato variety NUN00309 TOP by analyzing the seed coat or another maternal tissue of saidseed.

Using methods known in the art such as “reverse synthesis of breedinglines” or “reverse breeding,” it is possible to produce parental linesfor a hybrid plant such as tomato variety NUN 00309 TOP. A skilledperson can take any individual heterozygous plant (called a“phenotypically superior plant” in Example 2 of US 2015/0245570, whichis hereby incorporated by reference in its entirety; tomato variety NUN00309 TOP is such plant) and generate a combination of parental lines(reverse breeding parental lines) that, when crossed, produce thevariety NUN 00309 TOP. It is not necessary that the reverse breedingparental lines are identical to the original parental lines. Such newbreeding methods are based on the segregation of individual alleles inthe spores produced by a desired plant and/or in the progeny derivedfrom the self-pollination of that desired plant, and on the subsequentidentification of suitable progeny plants in one generation, or in alimited number of inbred cycles. Such a method is known from US2015/0245570 or from Wijnker et al., Nature Protocols Volume: 9, Pages:761-772 (2014) DOI: doi:10.1038/nprot.2014.049. Thus, the disclosureprovides a method for producing parental lines for a hybrid organism(e.g., tomato variety NUN 00309 TOP), comprising in one aspect: a)defining a set of genetic markers present in a heterozygous form (H) ina partially heterozygous starting organism; b) producing doubled haploidlines from spores of the starting organism; c) geneticallycharacterizing the doubled haploid lines thus obtained for the said setof genetic markers to determine whether they are present in a firsthomozygous form (A) or in a second homozygous form (B); and d) selectingat least one pair of doubled haploid lines that have complementaryalleles for at least a subset of the genetic markers, wherein eachmember of the pair is suitable as a parental line for the hybridorganism.

In another aspect, the method for producing parental lines of a hybridorganisms, e.g., of tomato variety NUN 00309 TOP, which when crossedreconstitute the genome of tomato variety NUN 00309 TOP, comprising:

-   -   a) defining a set of genetic markers that are present in a        heterozygous form (H) in a partially heterozygous starting        organism;    -   b) producing at least one further generation from the starting        organism by self-pollination (e.g., F2 or F3 generation);    -   c) selecting at least one pair of progeny organisms in which at        least one genetic marker from the set is present in a        complementary homozygous form (B vs. A, or A vs. B); and    -   d) optionally repeating steps b) and c) until at least one pair        of progeny organisms that have complementary alleles for at        least a subset of the genetic markers has been selected as        parental lines for a hybrid.

Thus, in one aspect, the disclosure relates to a method of producing acombination of parental lines of a plant of variety NUN 00309 TOP,comprising making doubled haploid cells from haploid cells or seed ofthat plant; and optionally crossing these parental lines to produce andcollecting seeds. In another aspect, the disclosure relates to acombination of parental lines produced by this method. In still anotheraspect, the combination of parental lines can be used to produce a seedor plant of variety NUN 00309 TOP when these parental lines are crossed.In still another aspect, the disclosure relates to a combination ofparental lines from which a seed or plant having all physiologicaland/or morphological characteristics of tomato variety NUN 00309 TOP(when the characteristics are determined at the 5% significance levelfor plants grown under the same conditions).

The disclosure also provides a method for producing parental lines forhybrid NUN 00309 TOP comprising: genetically characterizing a doubledhaploid line from tomato variety NUN 00309 TOP to determine whether oneor more genetic markers are present in a first homozygous form or in asecond homozygous form in said line, wherein the one or more geneticmarkers are present in a heterozygous form in tomato variety NUN 00309TOP; and selecting at least one pair of doubled haploid lines that havecomplementary alleles for the one or more the genetic markers, whereineach member of the pair is suitable as a parental line for a hybridorganism, optionally this method further comprises defining a set ofgenetic markers present in a heterozygous form in tomato variety NUN00309 TOP; and producing doubled haploid lines from tomato variety NUN00309 TOP. Doubled haploid lines generated as described herein can beused in such a method.

The disclosure also provides a combination of parental lines which, whencrossed, produce a seed or plant having all physiological and/ormorphological characteristics of tomato variety NUN 00309 TOP but one,two, or three which are different (when grown under the sameenvironmental conditions), as well as a seed or plant having allphysiological and/or morphological characteristics of tomato variety NUN00309 TOP but one, two or three which are different (when thecharacteristics are determined at the 5% significance level for plantsgrown under the same conditions).

In another aspect, a combination of a male and a female parental line oftomato variety NUN 00309 TOP can be generated by methods describedherein, for example, through reverse synthesis of breeding lines.

In another aspect, the disclosure provides a method of determining thegenotype of a plant described herein comprising detecting in the genome(e.g., a sample of nucleic acids) of the plant at least a firstpolymorphism or an allele. The skilled person is familiar with manysuitable methods of genotyping, detecting a polymorphism or detecting anallele including SNP (Single Nucleotide Polymorphism) genotyping,restriction fragment length polymorphism identification (RFLP) ofgenomic DNA, random amplified polymorphic detection (RAPD) of genomicDNA, amplified fragment length polymorphism detection (AFLP), polymerasechain reaction (PCR), DNA sequencing, allele specific oligonucleotide(ASO) probes, and hybridization to DNA microarrays or beads.Alternatively, the entire genome could be sequenced. The method may, incertain aspects, comprise detecting a plurality of polymorphisms in thegenome of the plant, for example by obtaining a sample of nucleic acidfrom a plant and detecting in said nucleic acids a plurality ofpolymorphisms. The method may further comprise storing the results ofthe step of detecting the plurality of polymorphisms on a computerreadable medium.

Also provided is a plant part obtainable from variety NUN 00309 TOP orfrom progeny of said variety or from a plant having all but one, two orthree tomato variety NUN 00309 TOP, or from a vegetatively propagatedplant of variety NUN 00309 TOP (or from its progeny or from a planthaving all or all but one, two or three physiological and/ormorphological characteristics which are different from those of tomatovariety NUN 00309 TOP, wherein the plant part is a fruit, a harvestedfruit, a part of a fruit, a leaf, a part of a leaf, pollen, an ovule, acell, a petiole, a shoot or a part thereof, a stem or a part thereof, aroot or a part thereof, a root tip, a cutting, a seed, a part of a seed,seed-coat or another maternal tissue which is part of a seed grown ontomato variety NUN 00309 TOP, or a hypocotyl, a cotyledon, a scion, astock, a rootstock, a pistil, an anther, or a flower or a part thereof.

A part of the plant of variety NUN 00309 TOP (or of progeny of saidvariety or of a plant having all physiological and/or morphologicalcharacteristics but one, two or three which are different from those ofsaid variety) encompasses any cells, tissues, organs obtainable from theseedlings or plants, such as but not limited to: a tomato fruit or apart thereof, a cutting, a hypocotyl, a cotyledon, seed coat, or pollen.

Such a plant part of variety NUN 00309 TOP can be stored and/orprocessed further. The disclosure thus also provides for a food or feedproduct comprising one or more of such parts from variety NUN 00309 TOPor from progeny of said variety, or from a derived variety, such as aplant having all but one, two or three physiological and/ormorphological characteristics of tomato variety NUN 00309 TOP.Preferably, the plant part is a tomato fruit or part thereof and/or anextract from a fruit or another plant part described herein comprisingat least one cell of tomato variety NUN 00309 TOP. The food or feedproduct may be fresh or processed, e.g., dried, grinded, powdered,pickled, chopped, cooked, roasted, in a sauce, in a sandwich, pasted,pureed or concentrated, juiced, pickled, canned, steamed, boiled, fried,blanched and/or frozen, etc.

In another aspect, the disclosure provides for a tomato fruit of varietyNUN 00309 TOP or a part of a fruit of said variety. The fruit can be inany stage of maturity, for example, immature or mature. In anotheraspect, the disclosure provides for a container comprising or consistingof a plurality of harvested tomato fruits or parts of fruits of saidvariety, or fruits of progeny thereof, or fruits of a derived variety.

Marketable tomato fruits are generally sorted by size and quality afterharvest. Alternatively, the tomato fruits can be sorted by expectedshelf life, pH, or Brix.

In another aspect, the plant, plant part or seed of tomato variety NUN00309 TOP is inside a container, for example, containers such as cans,boxes, crates, bags, cartons, Modified Atmosphere Packaging, films(e.g., biodegradable films), etc. comprising a plant or a part of aplant (fresh and/or processed) or a seed of tomato variety NUN 00309TOP. In a particular aspect, the container comprises a plurality ofseeds of tomato variety NUN 00309 TOP or a plurality of plant parts oftomato variety NUN 00309 TOP.

Tomato variety NUN 00309 TOP may also be grown for use as rootstocks(stocks) or scions. Typically, different types of tomatoes are graftedto enhance disease resistance, which is usually conferred by therootstock, while retaining the horticultural qualities usually conferredby the scion. It is not uncommon for grafting to occur betweencultivated tomato varieties and related tomato species. Methods ofgrafting and vegetative propagation are well-known in the art.

In another aspect, the disclosure provides to a plant comprising arootstock or scion of tomato variety NUN 00309 TOP.

All documents (e.g., patent publications) are herein incorporated byreference in their entirety, including the following cited references:

-   Naktuinbow Calibration book for Tomato (Lycopersicon esculentum    Mill), 2010-   UPOV, Guidelines for the Conduct of Tests for Distinctness,    Uniformity and Stability, TG/13/11, world-wide web at upov.int under    edocs/tgdocs/en/tg044.pdf.-   US Department of Agriculture, Agricultural Marketing Service,    Objective Description of Variety Tomato (Solanum lycopersicum or    Lycopersicon esculentum Mill), world wide web at    ams.usda.gov/services/plant-variety-protection/pvpo-c-forms, under    tomato.-   Bhatia, P., et al., Tissue Culture Studies of Tomato (Lycopersicum    esculentum), Plant Cell, Tissue and Organ Culture, 2004, vol. 78,    pp. 1-21.-   Hartz, et. al., “Processing Tomato Production in California,”    University of California Division of Agriculture and Natural    Resources, 2008, Publication 7228, pp. 1-5.-   Ince, A. G., et al., Genetic Relationship Within and Between    Capsicum Species, Biochem Genet, 2010, vol. 48, pp. 83-95.-   Needleman, S. B., et. al., A General Method Applicable to the Search    for Similarities in the Amino Acid Sequence of Two Proteins, Journal    of Molecular Biology, 1970, vol. 48(3), pp. 443-53.-   Nikolova, V., et. al., Diploidization of Cucumber (Cucumis sativus    L.) Haploids by Colchini Treatment, Acta Societas Botanicorum    Poloniae, 1996, vol. 65, pp. 311-317.-   Pisanu, A. B., et. al., “Yield and Biometric Characteristics of 9    Clones Selected from the Population of “Spinoso sardo” Artichokes,    Acta Hort., 2004, ISHS 660, pp. 83-89.-   Rice et al., EMBOSS: The European Molecular Biology Open Software    Suite, Trends in Genetics, 2000, vol. 16, Issue 6. pp. 276-277.-   Sharifova, S., et. al., “Assessment of Genetic Diversity in    Cultivated Tomato (Solanum lycopersicum L.) Genotypes Using RAPD    Primers”, Journal of Horticultural Research, 2013, vol. 21, no. 1,    pp. 83-89.-   Strange, et. al., “Fresh-Market Tomato Production in California,”    University of California Division of Agriculture and Natural    Resources, 2000, Publication 8017, pp. 1-8.-   Vidavsky F, Czosnek H., 1998, Tomato Breeding Lines resistant and    tolerant to tomato yellow leaf curl virus issued from Lycopersicum    hirsutum, Phytopathology, Sep; 88(9):910-4.-   Vos, P., et al., AFLP: A New Technique for DNA Fingerprinting 1995,    Nucleic Acids Research, 1995, vol. 23, No. 21, pp. 4407-4414.-   Wijnker, E., et al., Hybrid Recreation by Reverse breeding in    Arabidopsis thaliana, Nature Protocols, 2014, vol. 9, pp. 761-772.-   US2008/0222949-   U.S. Pat. No. 9,125,353-   US 2002/0010953-   U.S. Pat. No. 6,060,648-   EP 1057401-   EP 1428425-   US 2008/0222949-   US 2015/0126380-   US 2015/0245570    Development of Tomato Variety NUN 00309 TOP

The hybrid variety NUN 00309 TOP was developed from a male and femaleproprietary inbred line of Nunhems, selected mainly for its fruitquality, size, yield, and flavor. The female and male parents werecrossed to produce hybrid (F1) seeds of tomato variety NUN 00309 TOP.The seeds of tomato variety NUN 00309 TOP can be grown to produce hybridplants and parts thereof (e.g., tomato fruit). The hybrid variety NUN00309 TOP can be propagated by seeds or vegetatively.

The hybrid variety is uniform and genetically stable. This has beenestablished through evaluation of horticultural characteristics. Severalhybrid seed production events resulted in no observable deviation ingenetic stability. Coupled with the confirmation of genetic stability ofthe female and male parents the Applicant has concluded that tomatovariety NUN 00309 TOP is uniform and stable.

Deposit Information

A total of 2500 seeds of the hybrid variety NUN 00309 TOP was made andaccepted according to the Budapest Treaty by Nunhems B. V. on Aug. 10,2022 at the NCIMB Ltd., Ferguson Building, Craibstone Estate, Bucksburn,Aberdeen AB21 9YA, United Kingdom (NCIMB). The deposit has been assignedNCIMB number 44019. A statement indicating the viability of the samplehas been provided. A deposit of tomato variety NUN 00309 TOP and of themale and female parent line is also maintained at Nunhems B. V. The seedlot number for tomato variety NUN 00309 TOP is 30865701002.

The deposit will be maintained in NCIMB for a period of 30 years, or 5years after the most recent request, or for the enforceable life of thepatent whichever is longer and will be replaced if it ever becomesnonviable during that period. Access to the deposit will be availableduring the pendency of this application to persons determined by theDirector of the U.S. Patent Office to be entitled thereto upon request.All restrictions imposed by the depositor on the availability to thepublic of the deposited material will be irrevocably removed upon thegranting of the patent. Applicant does not waive any rights grantedunder this patent on this application or under the Plant VarietyProtection Act (7 U.S.C. § 2321 et seq.). Accordingly, the requirementsof 37 CFR § 1.801-1.809 will be satisfied.

Characteristics of Tomato Variety NUN 00309 TOP

The most similar variety to NUN 00309 TOP is AB 0311, a variety fromSeminis.

In Tables 1 and 2, a comparison between tomato variety NUN 00309 TOP andthe Reference Variety is shown based on a trial in Acampo, Calif., USA.Seeding date: Apr. 30, 2020; Transplanting date: Jun. 2, 2020;Harvesting date: Aug. 26, 2020. In Table 3, the distinguishingcharacteristics between tomato variety NUN 00309 TOP and the ReferenceVariety is shown.

A trial of 30 plants of each variety, from which at least 15 plants orplant parts were randomly selected and used to measure characteristics.For numerical characteristics averages were calculated. Fornon-numerical characteristics, the type/degree were determined.Similarity and differences between two different plant lines orvarieties can be determined by comparing the number of morphologicaland/or physiological characteristics (e.g., the characteristics aslisted in Tables 1-2) that are the same (i.e., statistically notsignificantly different) or that are different (i.e., statisticallysignificantly different) between the two plant lines or varieties whengrown under the same environmental conditions. A numericalcharacteristic is considered to be “the same” when the value for anumeric characteristic is not significantly different at the 1% (p<0.01)or 5% (p<0.05) significance level, using T-test, a standard method knownto the skilled person. A non-numerical or “degree” or “type”characteristic is considered “the same” when the values have the same“degree” or “type” when scored using USDA and/or UPOV descriptors, ifthe plants are grown under the same environmental conditions. In oneaspect, a statistical analysis using T-test at 5% significance level isprovided (see, Tables 4-14).

In one aspect, the disclosure provides a tomato plant having thephysiological and morphological characteristics of tomato variety NUN00309 TOP as presented in Tables 1-2, when grown under the sameenvironmental conditions.

TABLE 1 Characteristics of Tomato Variety NUN 00309 TOP and theReference Variety (USDA descriptors) Application Variety ReferenceVariety Characteristics (NUN 00309 TOP) (AB 0311) Mature Plant (atmaximum vegetative development): Plant growth: Determinate DeterminateIndeterminate, Determinate Form: Normal Normal Lax, open; Normal,Compact, Dwarf, Brachytic Size of canopy: Medium Medium Small, Medium,Large Plant habit: Semi-erect Semi-erect Sprawling, Semi-erect, ErectStem: Branching: Profuse Profuse Sparse, Intermediate, Profuse Branchingat cotyledonary or first leaf node: Present Present Present, Absent No.of nodes between 1^(st) inflorescence: Few Few Few (1-4), Several (5-9),Many (10 or more) No. of nodes between early (1^(st)-2^(nd,)2^(nd)-3^(rd)) Few Few inflorescence: Few (1-4), Several (5-9), Many (10or more) No. of nodes between later developing Few Few inflorescences:Few (1-4), Several (5-9), Many (10 or more) Pubescence on younger stems:Smooth (no long hairs) Smooth (no long hairs) Smooth, Sparsely hair,Moderately hairy, Densely hairy or wooly Leaf (mature leaf beneath the3^(rd) inflorescence) Morphology (type of leaf blade): BipinnateBipinnate Bipinnate, Pinnate Margin of major leaflets: Nearly entireShallowly toothed or Nearly entire, Shallowly toothed or scallopedscalloped, Deeply toothed or cut toward base Marginal rolling orwiltiness: Absent Slight Absent, Slight, Moderate, Strong Surface ofmajor leaflets: Smooth Smooth Smooth, Rugose Pubescence: Normal NormalSmooth, Normal, Hirsute, Wooly Inflorescence (3^(rd) inflorescence)Type: Simple Simple Simple, Forked, Compound Leafy or runninginflorescence: Occasional Occasional Absent, Occasional, Frequent Type(2^(nd) and 3^(rd) truss): Mainly uniparous Mainly uniparous Flower:Corolla color: Yellow Yellow Yellow, Old gold, White or tan Fruit(3^(rd) fruit of 2^(nd) or 3^(rd) cluster): Fruit shape: Pear Plum oregg shape Flattened, Slightly flattened, Round, High Round, Pear,Lengthened-pear, Plum or egg shape, Heart, Lengthened cylindrical,Angular Shape of transverse section: Round Round Round, Flattened,Angular, Irregular Shape of blossom end: Flat Flat Indented, Flat,Nippled, Tapered Shape of stem end: Indented Indented Flat, IndentedShape of pistil scar: Dot Dot Dot, Stellate, Linear, IrregularAbscission layer: Absent (jointless) Absent (jointless) Present, AbsentPoint of detachment of fruit at harvest: At calyx attachment At calyxattachment At pedicel joint, At calyx attachment Diameter of fruit atwidest point, mm: 46.86 mm 49.61 mm Mature fruit weight, g: 78.0 g 85.07 g  No. of locules: Two to three Two to three Two to three, Threeor four, Five or more Fruit surface: Smooth Smooth Smooth, Slightlyrough, Moderately rough or ribbed Fruit base color at mature-greenstage: Yellow green Light green Light green, Yellow green, Apple ormedium green, Light gray-green, Dark green Fruit pattern at mature-greenstage: Uniform green Uniform green Uniform green, Green-shouldered,Radial stripes on sides of fruit Fruit color, full ripe: Red Red White,Yellow, Orange, Pink, Red, (RHS 172B) (RHS 172B) Brownish, Greenish, Redwith purple, Yellow with purple, Purple Flesh color, full ripe: Red RedYellow, Red/Crimson, Pink, Orange, (RHS 34A) (RHS 34A) Brownish,Greenish Flesh color: Uniform Uniform Uniform, With lighter and darkerareas in walls Locular gel color of table-ripe fruit: Red Red GreenmYellow, Red Fruit core: Present Present Coreless, Present Soluble solidsas Brix: 5.93% 5.82% Resistance: Meloidoygne incognita (Mi) IntermediateResistant Intermediate resistant Verticillium sp. (Va and Vd) Race 0Highly Resistant Highly resistant Fusarium oxysporum f. sp. HighlyResistant Highly resistant lycopersici Race 0 Fusarium oxysporum f. sp.Highly Resistant Highly resistant lycopersici Race 1 Fusarium oxysporumf. sp. Highly Resistant Absent lycopersici Race 2 Tomato Spotted WiltVirus Highly Resistant Highly resistant Pseudomonas syringae pv. tomatoAbsent Intermediate resistant

TABLE 2 Characteristics of Tomato Variety NUN 00309 TOP and theReference Variety (UPOV descriptors) Application Variety ReferenceVariety Characteristics (NUN 00309 TOP) (AB 0311) Leaf (mature leafbeneath the 3^(rd) inflorescence): Attitude in relation to main stem (inmiddle Semi-erect Semi-erect to horizontal third of plant): Semi-erect,Horizontal, Drooping Size of leaflets (in middle of leaf): Large MediumVery small, Small, Medium, Large, Very large Intensity of green color:Dark Dark Very light, Light, Medium, Dark, Very dark Glossiness: WeakWeak Weak, Medium, Strong Blistering: Weak Weak Weak, Medium, StrongSize of blisters: Small Small Small, Medium, Large Attitude of petioleof leaflet in relation to Semi-erect to horizontal Horizontal main axis:Semi-erect, Horizontal, Semi-drooping Structure: Open Open Color: Darkgreen Dark green (RHS NN137B) (RHS NN137B) Anthocyanin coloration ofnerves: Absent Absent Leaf length, cm: 25.91 cm 23.13 cm Width, cm:18.44 cm 16.57 cm Petiole width, mm: 4.76 mm 4.28 mm Flower: Color:Yellow Yellow Yellow, Orange Anther color: Yellow Yellow Peduncle:Peduncle length, mm: 22.76 mm 26.33 mm Peduncle width, mm: 3.79 mm 3.51mm Mature Fruit: Green shoulder (before maturity): Absent Absent Absent,Present Intensity of green color excluding shoulder Light Very light(before maturity): Very light, Light, Medium, Dark, Very dark Greenstripes (before maturity): Absent Absent Size: Medium Medium Very small,Very small to small, Small, Small to medium, Medium, Medium to large,Large, Large to very large, Very large Shape in longitudinal section:Elliptic Ovate Flattened, Oblate, Circular, Oblong, Cylindrical,Elliptic, Cordate, Ovate, Obovate, Pyriform, Obcordate Ribbing atpeduncle end: Weak Weak Absent or very weak, Weak, Medium, Strong, Verystrong Depression at peduncle end: Weak Weak Absent or very weak, Weak,Medium, Strong, Very strong Size of peduncle scar: Medium Medium tolarge Very small, Small, Medium, Large, Very large Size of blossom scar:Small Small Very small, Small, Medium, Large, Very large Diameter ofcore in cross section in relation to Medium to large Medium totaldiameter: Cross section: Round Round Glossiness of skin: Medium MediumFirmness: Medium Medium Very soft, Soft, Medium, Firm, Very firm Fruitlength, mm: 66.99 mm 65.01 mm Thickness of pericarp, mm: 7.34 mm 7.16 mmStem scar diameter, mm: 6.90 mm 7.68 mm

TABLE 3 Distinguishing Characteristics between Tomato Variety NUN 00309TOP and the Reference Variety Application Variety Reference VarietyCharacteristics (NUN 00309 TOP ) (AB 0311) Leaf (mature leaf beneath the3^(rd) inflorescence) Margin of major leaflets: Nearly entire Shallowlytoothed or Nearly entire, Shallowly toothed or scalloped scalloped,Deeply toothed or cut toward base Marginal rolling or wiltiness: AbsentSlight Absent, Slight, Moderate, Strong Attitude in relation to mainstem (in Semi-erect Semi-erect to horizontal middle third of plant):Semi-erect, Horizontal, Drooping Size of leaflets (in middle of leaf):Large Medium Very small, Small, Medium, Large, Very large Attitude ofpetiole of leaflet in Semi-erect to horizontal Horizontal relation tomain axis: Semi-erect, Horizontal, Semi- drooping Leaf length, cm: 25.91cm 23.13 cm Width, cm: 18.44 cm 16.57 cm Petiole width, mm: 4.76 mm 4.28mm Peduncle: Peduncle length, mm: 22.76 mm 26.33 mm Fruit (3^(rd) fruitof 2^(nd) or 3^(rd) cluster): Fruit shape: Pear Plum or egg shapeFlattened, Slightly flattened, Round, High Round, Pear, Lengthened-pear,Plum or egg shape, Heart, Lengthened cylindrical, Angular Intensity ofgreen color excluding Light Very light shoulder (before maturity): Verylight, Light, Medium, Dark, Very dark Size of peduncle scar: MediumMedium to large Very small, Small, Medium, Large, Very large Diameter ofcore in cross section in Medium to large Medium relation to totaldiameter: Stem scar diameter, mm: 6.90 mm 7.68 mm Resistance: Fusariumoxysporum f. sp. Highly Resistant Absent lycopersici Race 2

The results of the T-test show significant differences between tomatovariety NUN 00309 TOP and the Reference Variety for mature leaf length,mature leaf width, petiole width, peduncle length, and stem scardiameter as shown in Tables 4-8.

Table 4 shows a significant difference between tomato variety NUN 00309TOP and the Reference Variety (p<0.01) on mature leaf length (cm) basedon the results of the trial conducted in the US during the trial season2020.

TABLE 4 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 23.30 20.30 Max.29.0 25.10 Median 25.60 23.70 Mean 25.91 23.13 Standard deviation 1.841.65

Table 5 shows a significant difference between tomato variety NUN 00309TOP and the Reference Variety (p=0.008) on mature leaf width (cm) basedon the results of the trial conducted in the US during the trial season2020.

TABLE 5 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 15.60 14.90 Max.22.50 21.20 Median 18.30 16.50 Mean 18.44 16.57 Standard deviation 1.981.56

Table 6 shows a significant difference between tomato variety NUN 00309TOP and the Reference Variety (p=0.005) on petiole width (mm) based onthe results of the trial conducted in the US during the trial season2020.

TABLE 6 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 4.11 3.45 Max. 5.474.96 Median 4.73 4.31 Mean 4.76 4.28 Standard deviation 0.43 0.43

Table 7 shows a significant difference between tomato variety NUN 00309TOP and the Reference Variety (p=0.002) on peduncle length (mm) based onthe results of the trial conducted in the US during the trial season2020.

TABLE 7 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 16.76 22.82 Max.28.53 31.07 Median 22.73 25.31 Mean 22.76 26.33 Standard deviation 2.912.69

Table 8 shows a significant difference between tomato variety NUN 00309TOP and the Reference Variety (p=0.05) on stem scar diameter (mm) basedon the results of the trial conducted in the US during the trial season2020.

TABLE 8 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 5.63 6.84 Max. 7.979.04 Median 6.92 7.59 Mean 6.90 7.68 Standard deviation 0.73 0.67

The results of the T-test show no significant differences between tomatovariety NUN 00309 TOP and the Reference Variety for peduncle width,mature fruit weight, mature fruit length, mature fruit diameter,thickness of pericarp, and % soluble solids as shown in Tables 9-14.

Table 9 shows no significant difference between tomato variety NUN 00309TOP and the Reference Variety (p=0.61) on peduncle width (mm) based onthe results of the trial conducted in the US during the trial season2020.

TABLE 9 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 3.09 3.10 Max. 4.364.68 Median 3.83 3.32 Mean 3.79 3.51 Standard deviation 0.39 0.41

Table 10 shows no significant difference between tomato variety NUN00309 TOP and the Reference Variety (p=0.389) on mature fruit weight (g)based on the results of the trial conducted in the US during the trialseason 2020.

TABLE 10 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 58.0 56.0 Max. 110.0140.0 Median 72.0 76.0 Mean 78.0 85.07 Standard deviation 17.82 25.69

Table 11 shows no significant difference between tomato variety NUN00309 TOP and the Reference Variety (p=0.398) on mature fruit length(mm) based on the results of the trial conducted in the US during thetrial season 2020.

TABLE 11 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 60.81 52.37 Max.75.06 77.47 Median 65.28 63.44 Mean 66.99 65.01 Standard deviation 4.997.42

Table 12 shows no significant difference between tomato variety NUN00309 TOP and the Reference Variety (p=0.100) on mature fruit diameter(mm) based on the results of the trial conducted in the US during thetrial season 2020.

TABLE 12 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 41.31 42.63 Max.55.26 57.43 Median 46.06 50.16 Mean 46.86 49.61 Standard deviation 4.374.47

Table 13 shows no significant difference between tomato variety NUN00309 TOP and the Reference Variety (p=0.554) on thickness of pericarp(mm) based on the results of the trial conducted in the US during thetrial season 2020.

TABLE 13 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 5.61 5.87 Max. 8.918.48 Median 7.15 7.04 Mean 7.34 7.16 Standard deviation 0.86 0.76

Table 14 shows no significant difference between tomato variety NUN00309 TOP and the Reference Variety (p=0.536) on % soluble solids basedon the results of the trial conducted in the US during the trial season2020.

TABLE 14 Application Variety Reference Variety Statistical Parameters(NUN 31812 WMW) (AB 0311) No. of samples 15 15 Min. 4.90 5.20 Max. 6.806.70 Median 5.70 5.80 Mean 5.93 5.82 Standard deviation 0.51 0.41

The invention claimed is:
 1. A plant, plant part, or seed of tomatovariety NUN 00309 TOP, wherein a representative sample of seed of saidvariety is deposited under Accession Number NCIMB
 44019. 2. The plantpart of claim 1, wherein said plant part is a fruit, a leaf, pollen, anovule, a cell, a scion, a root, a rootstock, a cutting, or a flower. 3.A seed that produces the plant of claim
 1. 4. A plant produced bygrowing the seed of claim
 1. 5. A tomato plant or a part thereof havingall of the physiological and morphological characteristics of the plantof claim 1, when grown under the same environmental conditions.
 6. Atissue or cell culture comprising regenerable cells of the plant orplant part of claim
 1. 7. The tissue or cell culture according to claim6, comprising cells or protoplasts from a plant part suitable forvegetative reproduction, wherein the plant part is a meristem, acotyledon, a hypocotyl, pollen, a leaf, an anther, a root, a root tip, apistil, a petiole, a flower, a fruit, a stem, or a stalk.
 8. A tomatoplant regenerated from the tissue or cell culture of claim 6, whereinthe plant has all of the physiological and morphological characteristicsof the plant of variety NUN 00309 TOP, when grown under the sameenvironmental conditions, and wherein a representative sample of seed ofsaid tomato variety is deposited under Accession Number NCIMB
 44019. 9.A method of producing the plant of claim 1 or a part thereof, saidmethod comprising vegetative propagating at least a part of the plant ofvariety NUN 00309 TOP, wherein a representative sample of seed of saidvariety is deposited under Accession Number NCIMB
 44019. 10. The methodof claim 9, wherein said vegetative propagating comprises regenerating awhole plant from a part of the plant of variety NUN 00309 TOP, wherein arepresentative sample of seed of said tomato variety is deposited underAccession Number NCIMB
 44019. 11. The method of claim 9, wherein saidpart is a cutting, a cell culture, or a tissue culture.
 12. Avegetatively propagated plant, or a part thereof, produced by the methodof claim 9, wherein the vegetatively propagated plant and part thereofhave all of the physiological and morphological characteristics of theplant of tomato variety NUN 00309 TOP, when grown under the sameenvironmental conditions, and wherein a representative sample of seed ofsaid tomato variety is deposited under Accession Number NCIMB
 44019. 13.A method of producing a tomato plant, said method comprising crossingthe plant of claim 1 with a second tomato plant one or more times,selecting a progeny tomato plant from said crossing and optionallyallowing the progeny tomato plant to form seed.
 14. A method ofproducing a tomato plant, said method comprising selfing the plant ofclaim 1 one or more times, [and] selecting a progeny tomato plant fromsaid selfing, and optionally allowing the progeny tomato plant to formseed.
 15. A method of introducing a desired trait into the plant ofclaim 1, said method comprising transforming the plant of claim 1 with atransgene that confers the desired trait, wherein the desired trait ismale sterility, herbicide tolerance, insect resistance, pest resistance,disease resistance, environmental stress tolerance, modifiedcarbohydrate metabolism, or modified protein metabolism.
 16. A tomatoplant produced by the method of claim 15, wherein the transformed plantcontains the desired trait and otherwise retains all of thephysiological and morphological characteristics of tomato variety NUN00309 TOP.
 17. A method of producing doubled haploid cells of the plantof claim 1, said method comprising making doubled haploid cells fromhaploid cells from the plant or plant part of tomato variety NUN 00309TOP, wherein a representative sample of seed of said tomato variety isdeposited under Accession Number NCIMB
 44019. 18. A plant comprising thescion or rootstock of claim
 2. 19. A container comprising the plant,plant part, or seed of claim
 1. 20. A method of producing a tomatofruit, said method comprising growing the plant of claim 1 until it setsat least one fruit, and collecting the fruit.
 21. A fruit produced bythe method of claim
 20. 22. A method of producing a modified tomatoplant having a single trait, said method comprises mutating a tomatoplant or plant part of variety NUN 00309 TOP, wherein a representativesample of seed of said tomato variety is deposited under AccessionNumber 44019, and wherein the modified plant has the single trait andotherwise all of the physiological and morphological characteristics ofthe plant of tomato variety NUN 00309 TOP, and wherein the trait is malesterility, herbicide tolerance, pest resistance, environmental stressresistance, modified carbohydrate metabolism, or modified proteinmetabolism.
 23. A method of determining the genotype of the plant ofclaim 1, said method comprising obtaining a sample of nucleic acids fromsaid plant and detecting in said nucleic acids a plurality ofpolymorphisms, thereby determining the genotype of the plant, andstoring the results of detecting the plurality of polymorphisms on acomputer readable medium.
 24. A method of producing a tomato seed, saidmethod comprising crossing tomato plants and harvesting the resultantseed, wherein at least one tomato plant is the plant of claim 1, andwherein a representative sample of seed of tomato variety NUN 00309 TOPis deposited under Accession Number NCIMB
 44019. 25. A method ofintroducing a single locus conversion into the plant of claim 1,comprising: a. crossing the plant of claim 1 with a second tomato plantcomprising a desired single locus to produce F1 progeny plants; b.selecting F1 progeny plants that have the single locus to produceselected F1 progeny plants; c. crossing selected F1 progeny plants withtomato variety NUN 00309 TOP to produce backcross progeny plants; d.selecting backcross progeny plants that have the single locus conversionand otherwise comprise all of the physiological and morphologicalcharacteristics of the tomato variety NUN 00309 TOP to produce selectedbackcross progeny plants; and e. repeating steps (c) and (d) one or moretimes in succession to produce selected second or higher backcrossprogeny plants that comprise the single locus conversion and otherwisecomprise all of the physiological and morphological characteristics ofthe tomato variety NUN 00309 TOP, wherein a representative sample ofseed of said tomato variety has been deposited under Accession NumberNCIMB
 44019. 26. The method of claim 25, wherein the single locusconversion confers male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, environmental stresstolerance, modified carbohydrate metabolism, or modified proteinmetabolism.
 27. A tomato plant produced by the method of claim 25,wherein the single locus converted plant comprises the single locusconversion and otherwise has all of the physiological and morphologicalcharacteristics of the plant of tomato variety NUN 00309 TOP.
 28. Amethod of producing a tomato plant derived from the plant of claim 1,comprising: a. preparing a progeny tomato plant derived from tomatovariety NUN 00309 TOP by crossing the plant [of any of the varieties] ofclaim 1 with itself or with a second tomato plant; b. crossing theprogeny plant with itself or a second tomato plant to produce seed of aprogeny plant of a subsequent generation; c. growing a progeny plant ofthe subsequent generation from said seed and crossing the progeny plantof the subsequent generation with itself or a second tomato plant; andd. repeating steps (b) and (c) for at least one more generation toproduce a tomato plant derived from tomato variety NUN 00309 TOP.